Fig 1: FUNDC2 regulates SLC25A11 protein. (A) Immunoblot analysis of FUNDC2, SLC25A11, SLC25A10, TIM23, TOM20, GPX4, and actin in WT and FUNDC2-KO MEF cells treated with 5 µM erastin or DMSO for 6 h. (B) Immunoblot analysis of FUNDC2, SLC25A11, SLC25A10, TIM23, TOM20, GPX4, and GAPDH in myocardium of WT and FUNDC2-KO mice treated with DOX or saline (control) for 4 d (n = 3 mice). (C) WT and FUNDC2-KO MEF cells treated with 5 µM erastin or DMSO for 3 h were collected for IP with lgG or anti-FUNDC2 antibodies and analyzed with anti-FUNDC2 or SLC25A11 antibodies, respectively. IB, immunoblot. IgG was used as the negative control. (D) Cardiac tissues of WT and FUNDC2-KO mice treated with DOX or saline (control) for 3 d were collected for IP with lgG or anti-FUNDC2 antibodies and analyzed with FUNDC2 or SLC25A11 antibodies, respectively. IgG was used as the negative control. (E) Dimeric and monomeric forms of SLC25A11 were analyzed in WT and FUNDC2-KO MEF cells and mitochondrial fractions treated with 5 µM erastin or DMSO for 6 h. Mitochondria fractions isolated from the cells and total cellular lysates with the presence or absence of SDS were separated by native PAGE, blotted to NC membranes, and incubated with antibodies against SLC25A11, FUNDC2, VDAC1, and actin. (F) Dimeric and monomeric forms of SLC25A11 were analyzed in the cardiac tissues and mitochondrial fractions from WT and FUNDC2-KO mice treated with DOX or saline (control) for 4 d. Mitochondria fractions isolated from cardiac tissues and tissue lysates with the presence or absence of SDS were separated by native-PAGE, blotted to NC membrane, and incubated with antibodies against SLC25A11, FUNDC2, VDAC1, and GAPDH. Data in A and C–F were obtained from three independent experiments. Statistical significance was calculated by Student’s t test (C and D) and one-way ANOVA (A, B, E, and F); *P < 0.05, **P < 0.01, ***P < 0.001 and ns indicates no significance.