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. 2020 Oct 22;8:585879. doi: 10.3389/fcell.2020.585879

FIGURE 1.

FIGURE 1

RRAD knockout did not affect the pluripotency nature of hESC and the ability of cardiac differentiation. (A) Schematic of human RRAD locus and editing site of designed gRNA-epiCRISPR/Cas9. (B) Sequencing data shows deletion of 55 bases in RRAD knockout hESC lines. (C) Immunostaining for the pluripotency markers SSEA4 and OCT4 of RRAD knockout hESC lines. Scalebar, 50 μm. (D) Schematic of cardiac differentiation using standard small molecule-based protocols. (E) Western blot shows RAD expression in WT and KO CMs at days 15. (F,G) Flow Cytometry for TNNT2 staining in representative WT and KO CMs at days 10 without purification. (H,I) Immunostaining for protein expression of MLC2v and MLC2a in WT and KO CMs at days 30. Scalebar, 50 μm. Data are expressed as means ± S.E.M. of three independent experiments. ns, not significant.