Fig. 2

Characterization of putative LSC marker expression during hPSC-LSC differentiation. a Representative morphology and protein expression of the cultures at selected time points. Scale bars, 100 μm for all images in the same column. Cell nuclei counterstained with DAPI (blue). b Marker expression differences in the d10 and d24 populations. Five images per sample and a minimum of 1400 cells per time point were analyzed for each marker from cytospin samples. c Representative IF image of ∆Np63 and p63α double-staining in a d24 cytospin sample. Scale bar, 100 μm for both c and d. d Representative IF image of ABCG2 and p63α double-staining in a d10 cytospin sample. e p63α and ABCG2 expression in d10 and d24 hPSC-LSCs. Five images per sample and a minimum of 3 000 cells per time point were analyzed from cytospin samples. P > 0.05. f The level of ABCG2 protein expression in UD-hPSCs and in d10 and d24–26 hPSC-LSCs, analyzed with flow cytometry. g The ABCG2 mRNA expression levels in UD-hPSCs and in d10 and d24 hPSC-LSCs analyzed with qRT-PCR. All representative data are presented with the hESC line Regea08/017. All quantitative data are presented as the mean + SD, and n marks the individual cell differentiation batches serving as biological replicates. Statistical analyses were carried out using the Mann-Whitney U test. *P ≤ 0.05