Fig. 5
IGF-II overexpression inhibited PSC proliferation while promoting differentiation. pME 18s PSCs were stably transfected with IGF-II and/or shRNA to inhibit IGF-II, as indicated. a Western blot analysis was performed to detect IGF-II in cell derivatives from various clones at differentiation day 17. b Quantification of a (n = 6). IGF-II level was normalized to β-actin level and is presented in a logarithmic scale of the fold change relative to pME 18s D3-derived cardiomyocytes. c Cell numbers were counted at day 17 (n = 8). d–j Representative images of derivative live cultures at day 17 from pME 18s D3, pME 18s ESCs, pME 18s PSCs, pME 18s PSCs+IGF-II shRNA, pME 18s IGF-II-PSCs, pME 18s IGF-II-PSCs+control shRNA, and pME 18s IGF-II-PSCs+IGF-II shRNA, respectively. Scale bar, 50 μm. k–q Periodic acid-Schiff-stained images of cell derivatives of (d–j) on day 17. Scale bar, 1 mm. r [14C] phenylalanine incorporation analysis was performed to examine protein synthetic activity in cell derivatives from various derivatives on day 17 (n = 12). s MTT assay of cell derivatives on day 17 (n = 8). Data are presented as fold change relative to pME 18s D3-derived cells. *p < 0.05 vs. pME 18s PSCs; ∆p < 0.05 vs. pME 18s IGF-II-PSCs+control shRNA; #p < 0.05 vs. pME 18s D3 or pME 18s ESCs