Unsupervised hierarchical clustering of iHPCs induced by 9 or 10 TFs

To clarify the gene expression profile of iHep, microarray analysis was performed using iHeps induced by 10 TFs (Foxg1, Lcor, Hnf3b, Hnf4a, Foxo6, Cdx2, Tcf1, Foxa3 ,Tcf2, Onecut1) and 9 TFs (Onecut1 was omitted from 10 TFs). Unsupervised hierarchical clustering indicated that iHep is expressing a global transcriptional profile more similar to that of HPCs rather than that of NPCs, and suggested that TFs present in the pool acted as inducing TFs. HPC (HB1 and HNG2) were established from fetal liver (E13.5) of C57BL6J and STOCK Tg(Nanog-GFP, Puro)1 Yam, respectively. iHeps were induced from NPC (NSBAg2, established from an ES cell line BAg73C2 carrying beta-geo knock-in allele in Afp) using retroviral vectors (pMXs without drug-selection markers) of 9 or 10 transcription factors. Three weeks after the infection, G418 was added and cultured for 1 week before the harvest. NSBAg2 and NSEB5-2C were used for the data of NPC. GSM396240 and GSM336010 were used for the data of ESC.