In the current study, we detected the hypothetical proteins Cpn0146 0147 in the C. pneumo niae inclusion membrane and Cpn0284 0285 within the inclusion although all four were predicted to be Inc proteins. Furthermore, Cpn0146 0147 but not Cpn0284 0285 co localized with a host cell endoplas mic reticulum marker when expressed via transgenes sellckchem although the ER co localization did not significantly affect the subsequent C. pneumoniae infection. Results 1. Localization of Cpn0146 and 0147 in the inclusion membrane and Cpn0284 and Cpn0285 within the inclusion of C. pneumoniae infected cells Using antibodies raised with C. pneumoniae fusion pro teins, we detected the hypothetical proteins Cpn0146 and 0147 in the inclusion membrane while Cpn0284 and 0285 within the inclusion of C. pneumoniae infected Inhibitors,Modulators,Libraries cells.

Both pAb and mAb antibodies against either Cpn0146 or Cpn0147 consistently detected a dominant inclusion membrane signal similar to the signal Inhibitors,Modulators,Libraries revealed by the anti IncA, but not the anti CPAFcp, anti MOMP or anti HSP60 antibodies. We fur ther took Inhibitors,Modulators,Libraries advantage of the isotype difference in the light Inhibitors,Modulators,Libraries chains between the anti Cpn0147 mAb 7H10 and anti IncA mAb 2B12. 1 to co label these two proteins in the same samples and found that Cpn0147 and IncA partially overlapped with each other under both conventional fluorescence and confocal microscopes. Since IncA, encoded by the C. pneumoniae ORF cpn0186, is a known inclusion membrane protein, the above observations suggest that Cpn0146 and 0147 Inhibitors,Modulators,Libraries are also inclusion membrane proteins.

Interestingly, the antibodies raised with Cpn0284 and 0285 fusion proteins labeled dominant signals within the inclusions, similar full report but not identical to the signals revealed by the anti MOMP or anti HSP60 antibodies. It is worth noting that the anti Cpn0284 and 0285 antibodies only detected strong signals in small but not large inclusions while both the anti MOMP and anti HSP60 antibodies detected all inclusions regardless of size. Since the small inclusions are mainly full of RBs while large inclusions full of EBs under the experimental conditions, we can speculate that Cpn0284 and 0285 are likely to be RB specific proteins. 2. Specificity of the anti chlamydial fusion protein antibodies Due to the fact that chlamyial antigens can be picked up by nonspecific antibodies, we further used several approaches to confirm the antibody binding specificities. First, a Western blot assay was used to measure the reactiv ity between the anti fusion protein antibodies and the GST fusion proteins. The anti Cpn0146, 0147, 0284, 0285 0186 antibodies only recognized the corresponding fusion proteins without obvious cross reaction with each other despite the common GST tag shared by all fusion proteins.