FIG. 2.
RCCS impairs embryonic development of cKit+ cells in an iPSCkit-Cre model of cardiogenesis. (A, B) Quantification of spontaneously beating EBs (A) and spontaneously beating EBs expressing enhanced green fluorescent protein (EGFP) (B) under SC or RCCS (n = 3/group). (C, D) Live tissue imaging of EGFP and Discosoma sea anemone-derived red fluorescent protein (DSRED) in SC (C) and RCCS-grown (D) iPSCkit-Cre-derived EBs, following NC differentiation and induction of Cre-mediated recombination with 4-hydroxytamoxifen. (E–H) Confocal immunofluorescence showing colocalization of EGFP, NKX2.5, and cardiac troponin I in iPSCkit-Cre-derived cKit+ NCs, grown under SC. Two-tailed t-test, *P < 0.05, ***P < 0.0005. Values are mean ± SEM. Scale bars 150 μm (C, D) and 10 μm (E–H). EBs, embryoid bodies; iPSC, induced pluripotent stem cells; NC, neural crest.