Profiling Pluripotency Transcription Factors

To examine mRNA expression of the transcription factors SOX2, KLF4, cMYC, OCT4 and Nanog, total RNA was isolated from stromal cells (passage 1, n = 3 for each source) cultured in different media using High Pure RNA isolation kit (Roche Diagnostics, Rotkreuz, Switzerland) according to manufacturer’s protocol. cDNA synthesis was done as described previously [35 (link)]. qRT-PCR analysis was performed using a LightCycler 480 II and LightCycler 480 SYBR Green I Master reagent (both Roche Diagnostics) according to manufacturer’s instructions. For normalization of sample material, human Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used. Data analysis was done as described [36 (link)], heat maps were done using ClustVis tool [37 (link)]. For qRT-PCR primer sequences are according to Lee et al. [38 (link)]. For detailed sequence information see also Additional file 3.