fig5

Figure 5. TNNI3 p.R168Q mutation regulates FASN expression through EGFR. (A) PPI network of TNNI3 generated using STRING 11.0. Nodes represent proteins. Edges represent PPIs. (B) EGFR protein levels measured by Western blot were detected in Tnni3^+/+ and Tnni3^R186Q/R186Q mice ventricular myocardium (n = 3 per group). The results are expressed as the means ± SEM and a representative experiment is shown. *P < 0.05, **P < 0.01, ***P < 0.001. The statistical test was done with unpaired T-test. GAPDH was used as the loading control. (C) Western blot analysis of FASN and EGFR in NRCMs of the Ad-WT, Ad-Mut and Ad-Mut +gefitinib groups (n = 3 per group). GAPDH was used as the loading control. (D) Co-IP for endogenous cTnI and EGFR in cultured cardiomyocytes of the Ad-WT and Ad-Mut groups (n = 3 per group). (E) Direct interaction of EGFR with cTnI by GST-pull down assay of Ad-WT and Ad-Mut groups (n = 3 per group). (F) Direct interaction of EGFR with cTnI by CO-IP assay in Tnni3^+/+ and Tnni3^R186Q/R186Q mice (n = 3 per group). (G) Representative GST pull-down assays showing that GST-labelled WT-cTnI pulled down recombinant EGFR, but GST-labelled Mut-cTnI showing a decreased binding to EGFR. The results are expressed as the means ± SEM and a representative experiment is shown. *P < 0.05, **P < 0.01. The statistical test was done with unpaired T-test. For Western blot analysis, GAPDH was used as the loading control.