Each of the resulting 19 recombinant plasmids was then introduced into both the wild type (FJ1) and the phaR mutant (FJR1) of R. sphaeroides and analyzed for luciferase activity (Table 2). Results showed that the luciferase activity derived from the wild-type (FJ1) R. sphaeroides harboring recombinant plasmids that carried any of the ZD1839 concentration DNA fragment (FP1, FP1-5, FP1-6, FP1-12, FP1-13, FP1-14, FP1-15, FP1-16, and FP1-17) shown to be able to bind the PhaR protein was approximately 50% (ranging from 2.0 ± 0.1 to 2.3 ± 0.4 RLU) of those (ranging from 4.1 ± 0.3 to 4.8 ± 0.2 RLU) containing the mutated PhaR-binding site to which the PhaR protein could not bind. However, all of the

19 luxAB fusion constructs yielded similar levels of luciferase activity in the phaR mutant (FJRI) of R. sphaeroides. These results strongly suggest that PhaR represses phaP expression. We have previously found that the PhaR protein regulates the expression of the phaP gene, which encodes phasin in R.

Ku0059436 sphaeroides FJ1. While investigating how PhaR regulates phaP expression, we found two 11-bp motifs (CTGCGGC(T)GCAG) present in the promoter region of the phaP gene. Because extensive searches of the GenBank failed to detect the presence of this sequence in the genomes of other bacteria, we characterized the sequence and determined its nucleotide residues that are important for the binding of PhaR. Results showed that the spacer region of this motif was not critical and could be replaced by any three or four bases. However, any base deletion or substitution in the two dyad regions of the palindrome rendered the motif unable to bind PhaR. As oxyclozanide mentioned above, two copies of the PhaR-binding motif exist in the promoter region of phaP. Multiple copies of such motif are also found in

other bacteria. For example, six 18-bp motifs of TGTCACCAACGGGCACTA that have been shown to be the binding site of the PhaR protein of Azotobacter vinelandii are present in the phbR–phbB intergenic region of the organism (Peralta-Gil et al., 2002). Similarly, three PhaR-binding sites with the sequence GCAMMAAWTMMD, where M, W, and D represent A or C, A or G, and A, G, or C, respectively, are found in the promoter region of the phaP gene of Ralstonia eutropha (Potter et al., 2005). In Paracoccus denitrificans, two TGC-rich sequences (TGC1 and TGAII) in the promoter region of the phaP gene were identified as the PhaR-binding sequences (Kojima et al., 2006). The sequences of TGCI and TGCII are CTGCACCGCAGCAA and TGCAATGCTGCGGTGCAG, respectively. These two sequences are similar to the consensus PhaR-binding sequence (CTGCN3−4GCAG) of R. sphaeroides, which we have determined in this study. The significance of the existence of multiple copies of the PhaR-binding site in the genomes of various bacteria remains to be determined. The consensus PhaR-binding sequence (CTGCN3−4GCAG) of R.

To combine these two separate experimental data, event frequencies should be normalised by the unit length of the axon (axonal short-pause rates, axonal appearance and disappearance rates; see ‘Materials and methods’; Fig. 8). The axonal appearance and disappearance rates were measured from the same experimental

data shown in Fig. 3 (Fig. 1C). The short-pause rate of individual mitochondria was suppressed by TTX treatment at 3 weeks (Fig. 5B). However, the axonal short-pause rate was not changed by TTX treatment because the number of mobile mitochondria was increased by TTX treatment (Figs 3I and 8). By using these normalised rates, we could calculate the stabilisation rates at different conditions ([SPSS]; Fig. 8). The stabilisation rate Fluorouracil mouse near synapses ([SPSS]synaptic) declined significantly from 2 to 3 weeks (1.01 vs. 0.53%) and was modulated by TTX treatment. Because stabilisation rates away from synapses ([SPSS]non-synaptic) were less affected by culture periods and TTX treatment, regulation of the stabilisation rate near synapses is likely Obeticholic Acid ic50 to be the parameter that is important for the control of mitochondrial replacement along the axon. Although the axonal appearance rate of

mitochondria near synapses ([MSS]synaptic) was more than twofold higher at 2 weeks, this increase was counterbalanced by the comparable rate of disappearance ([SSM]synaptic). It is likely that there exists a mechanism that keeps the balance between [MSS] and [SSM], as these rates were maintained in parallel in all experimental conditions (Fig. 8). This regulation may be important to keep the density of both synaptic and non-synaptic mitochondria constant with time. We report here the dynamic properties of axonal mitochondria using live-cell imaging with multiple sampling frequencies ranging from seconds to days. High-frequency image sampling is necessary to trace the accurate positions of mobile mitochondria, transported by motor proteins with their velocity of 0.1–1.4 μm/s (De Vos & Sheetz, 2007; MacAskill & Kittler, 2010).

In turn, the probability of transitions between stationary and mobile states is low (a few events per hour within an image area; Fig. 8) and time-lapse imaging with longer durations is required. Here we performed time-lapse imaging with high (intervals of 3 s), intermediate (intervals of 30 min) O-methylated flavonoid and low (intervals of 1 day) frequencies. Our results demonstrated that mitochondrial dynamics on multiple time scales differ between developmental stages and are regulated by neuronal activity and proximity to synaptic sites. To understand the dynamics of axonal mitochondrial distribution, mitochondrial properties in mobile and stationary states, and the transition process between them should be examined (Fig. 1). Our analyses revealed that the properties of stationary mitochondria are highly regulated by neuronal maturation and activity.

To combine these two separate experimental data, event frequencies should be normalised by the unit length of the axon (axonal short-pause rates, axonal appearance and disappearance rates; see ‘Materials and methods’; Fig. 8). The axonal appearance and disappearance rates were measured from the same experimental

data shown in Fig. 3 (Fig. 1C). The short-pause rate of individual mitochondria was suppressed by TTX treatment at 3 weeks (Fig. 5B). However, the axonal short-pause rate was not changed by TTX treatment because the number of mobile mitochondria was increased by TTX treatment (Figs 3I and 8). By using these normalised rates, we could calculate the stabilisation rates at different conditions ([SPSS]; Fig. 8). The stabilisation rate this website near synapses ([SPSS]synaptic) declined significantly from 2 to 3 weeks (1.01 vs. 0.53%) and was modulated by TTX treatment. Because stabilisation rates away from synapses ([SPSS]non-synaptic) were less affected by culture periods and TTX treatment, regulation of the stabilisation rate near synapses is likely EPZ015666 in vivo to be the parameter that is important for the control of mitochondrial replacement along the axon. Although the axonal appearance rate of

mitochondria near synapses ([MSS]synaptic) was more than twofold higher at 2 weeks, this increase was counterbalanced by the comparable rate of disappearance ([SSM]synaptic). It is likely that there exists a mechanism that keeps the balance between [MSS] and [SSM], as these rates were maintained in parallel in all experimental conditions (Fig. 8). This regulation may be important to keep the density of both synaptic and non-synaptic mitochondria constant with time. We report here the dynamic properties of axonal mitochondria using live-cell imaging with multiple sampling frequencies ranging from seconds to days. High-frequency image sampling is necessary to trace the accurate positions of mobile mitochondria, transported by motor proteins with their velocity of 0.1–1.4 μm/s (De Vos & Sheetz, 2007; MacAskill & Kittler, 2010).

In turn, the probability of transitions between stationary and mobile states is low (a few events per hour within an image area; Fig. 8) and time-lapse imaging with longer durations is required. Here we performed time-lapse imaging with high (intervals of 3 s), intermediate (intervals of 30 min) L-NAME HCl and low (intervals of 1 day) frequencies. Our results demonstrated that mitochondrial dynamics on multiple time scales differ between developmental stages and are regulated by neuronal activity and proximity to synaptic sites. To understand the dynamics of axonal mitochondrial distribution, mitochondrial properties in mobile and stationary states, and the transition process between them should be examined (Fig. 1). Our analyses revealed that the properties of stationary mitochondria are highly regulated by neuronal maturation and activity.

Future studies should investigate the use of slower feedback update rates. Fourth, adjusting the relative contribution of attended and unattended pictures based on decoder output did not allow us to dissociate between the effect of neurofeedback and the effect of change in BOLD signal due to change in the perceptual input. Future neurofeedback designs should avoid changing object properties by using a more abstract neurofeedback such as adjusting the color of the background surrounding the hybrid picture depending on the results of the decoding. Finally, a decoder trained on separately presented pictures of faces and places might not be the optimal way of investigating

the effects of neurofeedback. This is because a decoder trained on faces and places will recruit only those regions that it finds useful for distinguishing between BIBW2992 price face and place pictures. Presenting decoder output as neurofeedback to the subjects may have little impact on their task performance because the regions that respond to neurofeedback may not be incorporated in the decoding model trained on just faces and places. Hence, even if the subject’s brain Panobinostat is responding to neurofeedback, the decoder may be unable to detect it. Therefore, it is necessary that future studies using MVPA-generated neurofeedback could aim to incorporate the brain regions

responsible for processing feedback into the model. In case of whole-brain decoding, nine regions were consistently used by the classifier

to drive the predictions. Among these regions was the left fusiform gyrus, which is usually associated with reading and word processing (McCandliss et al., 2003; Hillis et al., 2005; Dehaene & Cohen, 2011). However, this area has also been suggested to be sensitive to the conjunction of object and background scene information (Goh et al., 2004). This view is strengthened by invasive studies in primates that also pointed to the Tryptophan synthase presence of neurons in this area, which are responsive to the conjunction of object features (Baker et al., 2002; Brincat & Connor, 2004). The left fusiform gyrus may be showing more activity for place blocks than for face blocks because pictures of famous places in the stimulus set contained not only objects but also a wide variety of backgrounds. Pictures used in the face blocks rarely had objects in them. The right fusiform gyrus showed a preference for face blocks, whereas the left parahippocampal gyrus showed a preference for place blocks. These two regions have been implicated in many studies to be responsible for the processing of faces and places, respectively (Aguirre et al., 1996, 1998; Kanwisher et al., 1997; McCarthy et al., 1997; Epstein & Kanwisher, 1998). Furthermore, bilateral ligual gyri were also activated for place pictures. The lingual gyrus performs bottom-up perceptual analysis of a scene in order to recognize it.

Enterococcus faecalis is a robust bacterium and, to survive within GIT, Ivacaftor has to cope with various stresses such as acid pH, nutrient limitation and deleterious effects of bile. In addition, the ability of the cell to survive in a wide range of environments as well as its intrinsic resistance

to chemical and physical stresses and antibiotics favour E. faecalis prevalence in an over-medicated environment (Michaux et al., 2011). The capacity of E. faecalis to cause disease is based on the presence of some major virulence factors but is also fine-tuned by many subtle virulence/fitness factors. Several transcriptional regulators have already been shown to be correlated with virulence and stress response, e.g. Fsr, EtaRS, CylR, HypR, PerR, SigV and Ers (Qin et al., 2001; Gilmore et al., 2002; Teng et al., 2002; Verneuil et al., 2004, 2005; Riboulet-Bisson et al., 2008; Le Jeune et al., 2010a). We recently characterized the transcriptional regulator SlyA (Ef_3002) of E. faecalis. Using the Galleria mellonella model it has been shown that the

ΔslyA mutant is more virulent than the wild-type strain. PARP assay In addition, ΔslyA survives better in macrophages and has a greater ability to persist in organs of mice (liver and kidneys) (Michaux et al., 2011). DNA microarray experiments revealed that 117 genes were deregulated in the ΔslyA mutant compared to the parental strain, and that SlyA acts as a repressor and activator (Michaux et al., 2011). In this study, we attempt to find stress conditions that affect the transcription of slyA. Among several stresses tested corresponding to agents potentially encountered in GIT or during the infection process, we found that bile salts induced expression of slyA. Moreover, the growth of ΔslyA mutant was more affected in its growth when bile salts are present, in comparison Atazanavir with the parental strain. In addition, RT-qPCRs were performed to

identify new SlyA-regulated genes. The E. faecalis strains and plasmids used in this work are listed in Table 1. Routinely, cells were grown without shaking at 37 °C in M17 medium supplemented with 0.5% glucose (GM17). Growth of the wild-type, ΔslyA, and complemented strain was examined for several environmental variables. Overnight cultures were diluted in fresh GM17 to an OD600 nm of 0.1. Growth was examined under the following conditions: 0.08% bile salts, 2 mM H2O2, 2% ethanol, growth under agitation with glycerol as the sole energy source (which induces an intracellular oxidative stress; Bizzini et al., 2010), pH 5.5, elevated temperatures (45, 50 and 55 °C), 20 mg mL−1 lysozyme, and growth in horse serum and human urine. Cultures were incubated until an OD600 nm of 0.5 and harvested after 30 min of exposition to stresses mentioned above or after 3 h in serum or urine.

oxysporum’s 15 chromosomes have been acquired through HGT from a fungal source (Ma et al., 2010). One of these chromosomes (chromosome 14) is essential

for pathogenicity of tomato plants (Ma et al., 2010). Using a simple co-incubation procedure, the authors demonstrated that chromosome 14 could be transferred between different F. oxysporum’s strains converting nonpathogenic strains into a pathogenic strains (Ma et al., 2010). Initially, a large proportion of documented HGT events into fungi involved bacterial Navitoclax donors (Table 1). This phenomenon may be due to the fact that bacterial HGT events are easier to detect than eukaryotic transfers. Furthermore, the majority of systematic fungal genomic HGT searches performed to date have only searched for genes from a bacterial source (Hall et al., 2005; Fitzpatrick et al., 2008; Marcet-Houben & Gabaldon, 2010). Ignoring these experimental biases, there are a number of biological reasons why prokaryote to fungal HGT is more likely than eukaryotic to fungal HGT. First, eukaryotic genes contain introns, and incorrect

spicing of these could act as a barrier for eukaryotic to eukaryotic HGT (this may not be an issue between Selleckchem Cobimetinib closely related eukaryotes where intron structure and position are highly conserved (Stajich et al., 2007)). Secondly, the number and diversity of bacterial populations is considerably larger than that of eukaryotic populations; therefore, the pool of bacterial genes available in the environment is significantly larger (Keeling & Palmer, 2008). Another factor to be considered is the observation that bacteria contain operons of functionally related genes, meaning that the transfer of a relatively small segment of DNA from bacteria to fungi could result in the gain of a complete metabolic pathway. Whole metabolic pathway transfer from bacteria to fungi has yet to be discovered; however, a recent analysis reported that two of the six genes (BIO3 and BIO4) of the S. cerevisiae biotin pathway have been acquired through HGT from a bacterial source (Hall & Dietrich, 2007).

Recent analyses have Avelestat (AZD9668) started to locate fungal to fungal interspecies HGTs (Table 1). Interestingly, a number of these studies have uncovered evidence of horizontal transfer of entire metabolic pathways whose genes are clustered within the donor genome (Temporini & VanEtten, 2004; Khaldi et al., 2008; Mallet et al., 2010; Khaldi & Wolfe, 2011; Slot & Rokas, 2011). For example, Slot and Rokas recently showed that a ~57-kb genomic region containing all 23 genes of the sterigmatocystin (toxic secondary metabolite) pathway has been transferred from Aspergillus nidulans to Podospora anserina (Slot & Rokas, 2011). Very few incidences of eukaryote (nonfungal) to fungal HGT have been located; however, a recent phylogenomic analysis has located four plant to fungi transfers (Richards et al., 2009). Resolving the tree of life is a fundamental goal of biology.

, 2005). Clinical chemistry analyses were conducted on CHIR 99021 serum (ADVIA 1650, Bayer Healthcare Diagnostics). The following parameters were measured: alkaline phosphatase, aspartate transaminase (AST), alanine transaminase (ALT), creatine kinase, blood urea nitrogen, creatinine, bilirubin, albumin, total protein, serum iron, calcium, magnesium and glucose. CRP was analysed using a dendrimer-coupled cytidine diphosphocholine sandwich enzyme-linked immunosorbent assay

(ELISA) (Heegaard et al., 2009). Detection antibodies were from DAKO (Glostrup, Denmark) and pooled pig serum calibrated against a human CRP calibrator (DAKO A0073) was used as the standard. The detection limit was 67 ng mL−1 (human equivalents) and all samples were run in duplicate. IL-6 and IL-1β serum concentrations were determined by sandwich ELISAs from R&D Systems (Duoset DY686 and Duoset DY681, respectively; RG7422 ic50 Abingdon, UK). Samples were run in duplicate in a dilution of 1 : 2 with a detection limit of 125 pg mL−1 (IL-6) and 62.5 pg mL−1 (IL-1β), using R&D Systems calibrators as the standard. Tumour necrosis factor (TNF)-α serum concentrations were determined using a sandwich ELISA from R&D Systems (Quantikine PTA00). Samples were run in duplicate in a dilution of 1 : 2 with a detection limit of 46.8 pg mL−1, using R&D Systems calibrators as a standard. At inoculation,

most of the S. aureus-infected animals showed dyspnoea, which started about 1 min after the inoculation and lasted about 2 min. Two animals had apnoea and had to be ventilated mechanically by means of repeated pressure on

the thorax for about 2 min. The dyspnoea and apnoea were sometimes accompanied by repeated clonic seizures, each lasting approximately 5 s. After the respiration had become stable, diffuse erythema of the skin appeared in several of the pigs, but disappeared after GABA Receptor 10–15 min. Recovery from sedation was uneventful in all cases, and the animals were able to stand less than 1 h PI. Seven to eight hours PI, signs of clinical disease were observed in all the infected animals. They became lethargic and remained in lateral or sternal recumbency most of the time and stood up reluctantly on manipulation. The respiration was forced and the body temperature was elevated and remained high throughout the experiment. At 12 h PI, an acute abscess surrounded by a haemorrhagic rim was found in the lung of one S. aureus-infected animal (I-1). At 24 h, two of the infected animals (II-1 and II-3) had multiple haemorrhagic processes in the lungs. The third pig (II-2) had pulmonary oedema and hyperaemia as well as a single pulmonary abscess surrounded by a haemorrhagic rim. At 48 h, all infected animals had pulmonary processes, either in the form of petechiae or small abscesses.

The evidence-based medicine training that these pharmacists received appeared to have limited influence on OTC decision-making. More work could be conducted to ensure that an evidence-based approach is routinely implemented in practice. “
“The objective of our research was to compare the reported pharmacy sales RO4929097 datasheet of pseudoephedrine-based medication in state where the electronic recording of sales

is mandatory, Queensland, with a state where recording is voluntary, Victoria. Unidentified, unit-record, pseudoephedrine-based medication transaction data (known as ProjectSTOP), for both states, were made available by GuildLink Pty Ltd, the data custodians. Data provided dated from roll-out, 8 November 2005 (Queensland) and 10 August 2007 (Victoria) to 16 October 2012 (the last entry at the time of request). Data were stored on a secure, password-protected computer at the University of Queensland, Australia, where it was prepared and analysed. The rate of uptake of ProjectSTOP in Queensland compared with Victoria differed significantly; 1 year after roll-out, 72% of pharmacies in Queensland had used the system compared with 41% in Victoria. There were significant

differences in transaction rates between Queensland and Victoria; the transaction rate in Queensland was four times greater than Victoria 1 year after roll-out. Our data show that Victoria captured fewer cases of multiple purchases using the same identification (i.e. suspected pseudo-runner activity) than in Queensland (112 click here compared with 517 cases in 2011). Our findings show, not surprisingly, that by making the electronic recording of pseudoephedrine-based medication sales mandatory, there is increased uptake and Bupivacaine use of the recording system ProjectSTOP. Importantly, by using ProjectSTOP comprehensively,

the data can provide useful intelligence for the identification of trends and patterns of activity in relation to the diversion of pseudoephedrine-based medications. “
“This is the second of two papers that explore the use of mixed-methods research in pharmacy practice. This paper discusses the rationale, applications, limitations and challenges of conducting mixed-methods research. As with other research methods, the choice of mixed-methods should always be justified because not all research questions require a mixed-methods approach. Mixed-methods research is particularly suitable when one dataset may be inadequate in answering the research question, an explanation of initial results is required, generalizability of qualitative findings is desired or broader and deeper understanding of a research problem is necessary. Mixed-methods research has its own challenges and limitations, which should be considered carefully while designing the study. There is a need to improve the quality of reporting of mixed-methods research.

The electrode was first stabilized at zero oxygen consumption in fresh PDB with constant stirring in the thermo-balanced chamber at 30 °C before the fungal suspension was transferred to the chamber. Recordings of respiration rate were initiated after closing the chamber with an air-tight lid. At least 10 min

after initiating the recording of basal respiration, 4 μM of the uncoupler carbonyl cyanide m-chlorophenylhydrazone, 4 μM of the alternative oxidase (AOX) inhibitor salycil-hydroxamic acid (SHAM) and/or 4 μM of the complex III respiratory inhibitor see more antimycin A (AA), or 1 μM of the complex IV respiratory inhibitor potassium cyanide (KCN) were added to the chamber containing C. neoformans. Values are represented as the rate of O2 consumption in nanomoles min−1 ± SD. Statistical analysis was performed using prism version 5 (GraphPad Software). The results were compared by Student’s t-test or two-way anova test according to the data. In a previous study, we showed by absorbance readings (A595 nm) that 0.09 μM of microplusin inhibited 50% of the growth of C. neoformans (Silva et al., 2009). However, we did not determine whether

microplusin was fungicidal or fungistatic. We addressed this question by incubating C. neoformans Z VAD FMK (strain H99) with 10 μM microplusin. After 72 h incubation, the number of MP-treated yeast cells was 10-fold lower compared with non-MP treated cells (Fig. 1a). A similar result was obtained after 48-h incubation with microplusin (data not shown). To determine the viability of C. neoformans after exposure to MP, 100 yeast cells (MP-treated and non-MP treated systems) were plated onto Sabouraud agar medium. Although there was a trend toward a reduction in CFU after 48-h incubation in MP-treated cells, the CFU determinations were not significantly different (P-value = 0.1710) between the two culture conditions (Fig. 1b). Hence, microplusin predominantly has a fungistatic effect against C. neoformans. In addition, supplementation of PDB medium with 2.5 μM of CuCl2.6H2O Chloroambucil significantly impaired microplusin’s

activity against C. neoformans (Fig. 2). The protective effect of copper depended on the concentration of microplusin, as the inhibitory action of the compound was most pronounced at microplusin concentrations ≥1.56 μM. To test whether the copper depletion promoted by microplusin affected complex IV functioning, and therefore electron flow through classical respiratory pathway, we measured oxygen consumption of C. neoformans in the presence of different inhibitors of the electron transport complexes. In non-treated C. neoformans, electrons flow largely via the classical pathway, since inhibition of either complex III/cytochrome c reductase with 4 μM AA or complex IV/cytochrome oxidase with 1 μM KCN decreased oxygen consumption by ~70% (Fig. 3).

, 1981; Valladares et al., 2007). The potential localization of the oxygen-sensitive uptake hydrogenase to the honeycomb membrane structures could be of relevance for the function of

this enzyme, as one of the targets for the electrons released during H2 oxidation is the respiratory electron transport chain (Houchins & Burris, 1981), and because the respiration could lead to locally decreased levels of O2. However, because the fluorescence foci are also observed outside the polar regions, alternative explanations merit consideration. One such consideration is that the HupS–GFP is targeted for localized NVP-LDE225 degradation by proteases. An interesting reflection is that protease complexes that show similarities to the eukaryotic proteasome have been identified in some bacteria, i.e. the cyanobacterium Synechoccoccus and Bacillus subtilis (Kirstein et al., 2008; Simmons et al., 2008; Andersson et al., 2009). Furthermore, the protease clusters in B. subtilis are localized to the polar regions of the cells Cyclopamine and have been shown to be colocalized with protein aggregates (inclusion bodies) (Kirstein et al., 2008). Protein inclusion bodies are most often formed

during high-level protein expression in biotechnological applications (Wang, 2009), but because of the apparent low solubility of HupS–GFP, it could not be excluded that it forms a protein aggregate. The observation of fluorescent clusters in SHG over time during increased expression of HupS–GFP could indicate some form of inclusions. Even though it has been observed that a GFP fusion protein was not fluorescent upon formation of inclusion bodies (Drew et al., 2001), another study has shown that inclusion bodies of GFP fusion proteins indeed can be fluorescent (Garcia-Fruitos et al., 2005). To investigate the possibility

of HupS–GFP CHIR-99021 in vitro inclusions, TEM was used to compare heterocysts isolated from N2-fixing cultures of WT and SHG and to search for structural differences indicating inclusion bodies (Fig. S2). No such differences could be observed, although this is difficult to determine due to the many structures within the heterocyst, i.e. membranes and cyanophycin granules. Because HupS–GFP required strong denaturing conditions to be extracted, whereas most degradation products could be extracted without detergents, indicating a more soluble form, it is likely that full-length HupS–GFP forms the clusters. In the present study, the in vivo localization of the uptake hydrogenase is determined for the first time, using a HupS–GFP fusion protein reporter, as solely localized to the heterocysts of N. punctiforme. The subcellular fluorescence in fully mature heterocysts is either homogeneously distributed or localized in clusters, which may be of relevance for the function of the uptake hydrogenase.