Fig. 5

Overexpression of wild-type JPH2 rescues ECC impairment in KO-CMs. (A) Schematic diagram of the overexpression vector backbone for lentiviral transfection. (B) Western blot analysis of JPH2 expression in KO-CMs transfected with different MOIs (10, 15, and 20). Full-length blots are presented in Figure S8. (C) Representative TEM images of the JMC structure in JPH2-OE-KO-CMs. CMs were transduced with lentivirus at two weeks post-differentiation, and phenotypes were assessed two weeks after transduction. The JMC structures are highlighted with yellow arrows. Scale bar=100nm. (D) Bar graph showing the distance between the PM and SR membrane in WT-CMs, KO-CMs and JPH2-OE-KO-CMs. (E) Scatter plot of ECCD in WT-CMs, KO-CMs and JPH2-OE-KO-CMs. To eliminate potential effects of lentiviral transfection, WT-CMs and KO-CMs were also transfected with an empty vector. (F) Bar graph showing the coefficient of variation of ECCD in WT-CMs, KO-CMs and JPH2-OE-KO-CMs. Results are presented as means ± S.E.M. of 6 independent experiments. ns, not significant, ***P < 0.001; ****P < 0.0001, unpaired two-sided Student’s t test. Abbreviations: WT, WT-CMs; KO, KO-CMs; OE10, JPH2 overexpression at MOI=10; OE15, JPH2 overexpression at MOI=15; OE20, JPH2 overexpression at MOI=20; Ctrl, Empty vector transfection at MOI=20; JPH2-OEs, JPH2-OE-KO-CMs.