Fig. 4: Disruption of mitochondrial ROS-derived MERCSs attenuates cell viability. | Nature Communications

Fig. 4: Disruption of mitochondrial ROS-derived MERCSs attenuates cell viability.

From: ER-mitochondria contacts mediate lipid radical transfer via RMDN3/PTPIP51 phosphorylation to reduce mitochondrial oxidative stress

Fig. 4: Disruption of mitochondrial ROS-derived MERCSs attenuates cell viability.The alternative text for this image may have been generated using AI.

a Rotenone or antimycin A treatment of RMDN3 and VAPB knockdown cells reduced cell viability. The HeLa cells were transfected with the indicated siRNAs for 5 days with or without rotenone (50 nM) and antimycin A (50 nM) for 2 days before cell viability was measured. Cell viability was determined and expressed as a fold change of si-NT. Viable cells were detected by cell viability assay using Cell Counting Kit-8. b The HeLa cells were transfected with the indicated siRNAs for 5 days with or without rotenone (50 nM) and antimycin A (50 nM) for 2 days before measuring cell viability. oxNAC (50 µM), NACS2 (50 µM) and mito-TEMPO (100 nM) were treated for 3 days before measuring cell viability. Cell viability was determined and expressed as a fold change of si-NT. Viable cells were detected by cell viability assay using Cell Counting Kit-8. c Lack of TPR domain does not rescue cell viability of RMDN3 knockdown with rotenone or antimycin A treatment. HeLa cells were transfected with RMDN3 siRNA for 5 days and with the indicated vectors for 3 days before measuring cell viability. Rotenone (50 nM) and antimycin A (50 nM) were treated for 2 days before cell viability was measured. Cell viability was determined and expressed as a fold change of si-NT. Viable cells were detected by cell viability assay using Cell Counting Kit-8. FLAG-resi-RMDN3 WT, FLAG-resi-RMDN3ΔFFAT, and FLAG-resi-RMDN3ΔTPR are RMDN3 RNAi-resistant vectors. d HeLa cells were transfected with RMDN3 siRNA for 5 days with or without rotenone (50 nM) and antimycin A (50 nM) for 2 days before measuring cell viability. The indicated inhibitors were treated for 2 days before measuring cell viability. Cell viability was determined and expressed as a fold change of si-NT. Viable cells were detected by cell viability assay using Cell Counting Kit-8. z-VAD-FMK (20 µM), necrostatin-1 (20 µM), ferrostatin-1 (Fer-1) (5 µM), and deferoxamine (DFO) (100 µM). Data are mean ± s.e.m. (n = 3, triplicate), and statistical significance was analyzed by one-way analysis of variance (ANOVA) (a-d). P values are indicated as ****p < 0.0001; n.s., not significant.

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