Fig. 3

RT-PCR analysis of HDDPCs collected through RNA-snMIFxC. (A) HDDPCs sorted. There are ~ 10 HDDPCs with weakly positive octamer-binding transcription factor 3/4 (OCT3/4) activity (a, defined as HDDPC-OCT3/4( +)L) and those with negative OCT3/4 activity (b, defined as HDDPC-OCT3/4(-)L). Similarly, there are ~ 10 HDDPCs with strongly positive OCT3/4 activity (c, defined as HDDPC-OCT3/4( +)S) and those with negative OCT3/4 activity (d, defined as HDDPC-OCT3/4(-)S). Bar = 100 μm. (B) RT-PCR analysis of HDDPC-sex determining region Y-box 2 (SOX2)( +), HDDPC-SOX2(-), HDDPC-OCT3/4( +)L, HDDPC-OCT3/4(-)L, HDDPC-OCT3/4( +)S and HDDPC-OCT3/4(-)S using primer sets for alkaline phosphatase (ALP), OCT3/4, SOX2 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). In the lower panel, comparison of the expression levels of each transcript between HDDPC-SOX2( +) and HDDPC-SOX2(-), HDDPC-OCT3/4( +)L and HDDPC-OCT3/4(-)L or HDDPC-OCT3/4( +)S and HDDPC-OCT3/4(-)S, after normalization, is shown. The intensity of each band in the left panel was determined using The ImageJ software. The level of each transcript was normalized to that of GAPDH mRNA, and the results obtained are represented as a graph. M, 100 bp ladder; -RT, PCR performed in the absence of reverse transcription products; HDDPC PC, RT-PCR products using cDNAs derived from unselected HDDPCs; iPSC PC, RT-PCR products using cDNAs derived from unselected HDDPC-derived induced pluripotent cells (iPSCs)