Fig. 4. Increased gene expression of hypertrophic signaling pathways in MLP-deficient hESC-CMs.

a Heatmap showing changes in the expression of genes involved in HCM signaling pathways, calcium handling, fibrosis, and autophagy in WT and MLP KO hESC-CMs at day 30. b qRT-PCR analysis of HCM related genes in WT and MLP KO CMs at day 30. c Heatmap showing the expression of HCM associated genes at days 15, 22, and 30 of cardiac differentiation. d Immunoblot analysis of HCM signaling (calcineurin A, CaMKIIδ, and phosphorylated CaMKII) and sarcomere proteins (MYH7, α-actinin, and cTnT) in WT and MLP KO hESC-CMs at day 30. e–j Quantification of calcineurin A, CaMKIIδ, P-CaMKII, MYH7, α-actinin, and cTnT normalized by GAPDH in WT and MLP KO hESC-CMs. Results are presented as means ± S.E.M. of three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant, unpaired two-sided Student’s t test