Extended Data Fig. 4: DNAJC15 does not affect MMP, TIM22 levels and protein turnover rates.
From: Stress adaptation of mitochondrial protein import by OMA1-mediated degradation of DNAJC15
a, Histogram plot of the TMRM fluorescence levels in wild-type (WT) HeLa cells after siRNA-mediated depletion of DNAJC15 (siJC15) or CCCP treatment for 2 h (50 µM). A representative experiment is shown (n = 3, biologically independent samples). Single cells were identified using a conventional FSC/SSC gating strategy, and the mean fluorescence intensity of the gated population was quantified. b, Quantification of the mean TMRM fluorescence intensity in (a). Data are means ± s.d. (n = 3, biologically independent samples). P values were calculated using a two-sided unpaired t-test. c, Stability of several mitochondrial proteins in wild-type (WT) HeLa cells depleted of DNAJC15 (siJC15) for 48 h after inhibition of cytosolic translation with emetine. A representative experiment is shown (n = 3, biologically independent samples). d, Degradation rates of the proteins in (c) normalized to time point 0 h. Data are means ± s.d. (n = 3, biologically independent samples). P values were calculated using a two-sided unpaired t-test. Quantile box plot show median (center line), 25th and 75th percentiles, whiskers show minimum and maximum values (1.5 * IQR distance from median, outliers not shown). e, Fraction of significantly changed mitochondrial proteins (FDR < 0.05) within all identified proteins of various mitochondrial subcompartments. f, Expression levels of TIM22 components and TIMM22 substrates in wild-type (WT) HeLa cells after siRNA-mediated depletion of DNAJC15 (siJC15) monitored by Western Blot. g, Quantification of the protein levels in (f). Data are means ± s.d. (n = 3, biologically independent samples). P values were calculated using a two-sided unpaired t-test. h, Boxplot visualizing the distribution of MitoCarta 3.0-annotated proteins according to their mitochondrial protein import in ref. 45. Quantile box plot show median (center line), 25th and 75th percentiles, whiskers show minimum and maximum values after outlier removal (1.5 * IQR distance from median, outliers not shown). Mann-Whitney U-test was performed (n = 5, biologically independent samples, including outliers).
