Fig. 1.

MARS2 regulates cellular redox state via p53. a. Cellular ROS level was analyzed by DCF-DA confocal microscopy and flow cytometry assay upon MARS2 knockdown in A549 non-small cell lung cancer (NSCLC) cells (n = 5). si-Cont indicates si-control RNA. b. Mitochondrial ROS level was analyzed by MitoSox confocal microscopy and flow cytometry assay upon MARS2 knockdown in A549 cells (n = 5). c. Cellular ROS level was analyzed by DCF-DA confocal microscopy and flow cytometry assay of A549 cells upon MARS2 knockdown and TP53 double knockdowns (n = 5). d. Western blot analysis of p53 protein level upon MARS2 knockdown and a rescue assay with exogenous MARS2 expression in A549 cells (n = 3). e. Western blot analysis of TIGAR protein level upon MARS2 knockdown and a rescue assay with exogenous MARS2 expression in A549 cells (n = 3). f. G6PDH activity assay of A549 cells upon MARS2 knockdown and double knockdowns of MARS2 and TP53 (n = 3). g. NADPH assay of A549 cells upon MARS2 knockdown and double knockdowns of MARS2 and TP53 (n = 4). h. ATP production profile of A549 NSCLC cells upon MARS2 knockdown was indicated by the ratio of glycolytic ATP production level, mitochondrial ATP production level (OXPHOS) and basal level (n = 5). i. ATP production profile of A549 NSCLC cells upon MARS2 and TP53 double knockdowns was indicated by the ratio of glycolytic ATP production level, mitochondrial ATP production level (OXPHOS) and basal level (n = 5). j. Extracellular acidification rate (ECAR) with MARS2 knockdown and double knockdowns of MARS2 + TP53 in A549 cells (n = 10). k. Oxygen consumption rate (OCR) with MARS2 knockdown and MARS2 + TP53 double knockdowns in A549 cells (n = 7). All the quantitative data in graphs are marked as the mean ± S.D from at least three independent samples. Statistical analyses of results were performed with Student's t-test or ANOVA followed by Tukey's test (*, P < 0.05, **, P < 0.01, ***, P < 0.001, #, P < 0.05 versus si-MARS2, ##, P < 0.01 versus si-MARS2).