Fig. 1

Generation and characterization of JPH2-knockout hESC-derived cardiomyocytes. (A) Schematic of the sgRNA targeting the MORN region 2 of JPH2, resulting in a 16-bp deletion from 333 amino acids. (B) Flow cytometry analysis of cardiomyocyte differentiation efficiency at day 15 post-differentiation. (C) Bar graph showing the percentage of TNNT2-positive cells in WT-CMs and KO-CMs. (D) Morphology of WT-CMs and KO-CMs at day 15 post-differentiation (40x magnification). (E) Western blot analysis confirming the knockout of JPH2 protein in KO-CMs. Full-length blots are presented in Figure S8. (F) Immunofluorescence staining of ventricular-specific marker MYL2 and atrial-specific marker MYL7 in WT-CMs and KO-CMs: MYL2 in green, MYL7 in red, DAPI in blue. Scale bar = 100 μm. Results are presented as means ± SEMs of 3 independent experiments. ns, not significant, unpaired 2-sided Student’s t test