For TIC experiments, animals were injected in the flank with OTBCs 86 L1 Ds Red cells diluted to 1, 50, 1, 000, 100, 000, and 1, 000, 000 cells in a 100 uL PBS Matrigel mixture. Tumor growth was monitored by caliper measurement and fluorescence imaging as described above. Gene expression microarrays A total of 11 cell lines were used for gene expression selleck chemicals Pazopanib analyses, 4 parental cell lines, 6 OTBCs, and 1 OCT4 siRNA cell line. In addition, one tumor sample generated from the OTBCs86 L1 cell line was used for gene expression ana lysis. From each sample, total RNA was purified, Inhibitors,Modulators,Libraries ampli fied, labeled, and hybridized by using Agilent 4 �� 44 K oligo microarrays. All microarray data have been deposited in the Gene Expression Omnibus under accession number GEO,GSE26539.

The probes Inhibitors,Modulators,Libraries genes were filtered by requiring the lowest normalized intensity values to be greater than 10 in both samples and controls. The normalized log2 ratios of probes mapping to the same gene were averaged to generate independent expression estimates. We also used available microarrays from the UNC337 dataset. For the UNC337 dataset, genes were med ian centered, and samples were standardized to zero mean and unit variance before other analyses were per formed. All microarray cluster analyses were displayed by using Java Treeview version 1. 1. 3. Average linkage hierarchical clustering was performed by using Cluster version 2. 12. Analysis of variance tests for gene expression data were performed using R.

OCT4 transduced Inhibitors,Modulators,Libraries breast cell gene signatures To build an OTBC signature, we first selected those genes that were significantly and differentially expressed between six OTBCs and their four respective parental cell lines by using two class paired SAMs and a less than 1% false discovery rate. The resulting upregulated and downregulated gene lists are shown in supplemental data. To estimate the expression of the OTBC signa ture across the intrinsic molecular subtypes of breast cancer, we calculated the mean expression of both gene lists in the entire med ian centered UNC337 dataset by using the subtype calls described in. Inhibitors,Modulators,Libraries Among the entire gene list Inhibitors,Modulators,Libraries of the OTBC signature, only three genes were found missing in the UNC337 dataset. thereby Immunofluorescence, flow cytometry, Western blot ting, immunohistochemistry, and small molecule epige netic inhibitor treatments are described in supplementary methods in Additional file 2. Primary and secondary antibodies were used in accordance with the recommendations of the manufacturer and are listed in Table S2 in Additional file 3.