qRT–PCR analysis of PKP2 reveals similar expression levels in wt and PKP2mut CMs (n = 3).
Left, Western blotting using an antibody directed against the N‐terminus of PKP2 demonstrates reduced expression of wild‐type PKP2 protein (PKP2 wt, 97 kDa) and absence of truncated A587fsX655‐PKP2 product (PKP2 mut, 72 kDa) in total lysates of PKP2mut CMs. Pan‐Cadherin is shown as a loading control. Right, densitometric readings for PKP2‐wt bands reveal an almost 50% reduction in PKP2mut myocytic lysates compared to wt ones. Values are expressed as the integrals (area × mean density) of each band normalized to Pan‐Cadherin and relative to wt; n = 3; *P < 0.01 vs. wt; t‐test.
Immunofluorescence analysis indicates an interrupted desmoplakin expression (DSP, red) at the plasma membrane of PKP2mut CMs compared to wt cells. cTNT (green) marks cardiomyocytes. Nuclei are stained with Hoechst 33258 (blue). Scale bars, 12.5 μm.
qRT–PCR analysis of pro‐apoptotic genes and white adipocytic markers shows similar expression levels in wt and PKP2mut CMs over time in culture in adipogenic medium. Brown/beige adipocytic markers were upregulated in PKP2mut compared to wt cells; n = 3; *P < 0.05, **P < 0.01 vs. wt CMs; t‐test.
Brown/beige adipocyte‐specific genes were similarly upregulated in PKP2mut CMs at 28 days in culture in adipogenic medium and wt iPSC‐derived brown/beige adipocytes at day 20 of differentiation compared to wt CMs as determined by qRT–PCR; n = 3; *P < 0.05, **P < 0.01, ***P < 0.001 vs. wt; t‐test.
qRT–PCR analysis of indicated genes shows similar activation in PKP2mut CMs and day‐20 iPSC‐derived brown/beige adipocytes after treatment with 1 mM 8‐Br‐cAMP for 48 h. Expression values are relative to basal conditions before treatment in each group; n = 3; *P < 0.05, **P < 0.01, ***P < 0.001 vs. corresponding PKP2mut CMs or brown/beige adipocytes at basal conditions; t‐test.
Data information: All data are shown as means ± SEM.
