Fig. 4: Loss of DDHD2 induces mitochondrial dysfunction in MEFs.

a DDHD2 KO MEFs were incubated with 2.5 µM CellROX Green for 30 min, and then stained with an antibody against Tom20 or cytochrome c and analyzed by IF microscopy. b WT and DDHD2 KO MEFs were incubated with 5 µM MitoSOX and 100 nM MitoTracker Green FM for 10 min, and then analyzed by IF microscopy without fixation. The fluorescence intensity of MitoSOX was quantified and is expressed as the ratio relative to that in WT MEFs. c The ATP contents in WT and DDHD2 KO MEFs were measured. The graph shows the ratio of the ATP level in DDHD2 KO MEFs to that in WT MEFs. d WT and DDHD2 KO MEFs were incubated with 2 µM JC-1 for 30 min, and then analyzed by IF microscopy without fixation. The ratio of Green/Red fluorescence intensity of JC-1 was determined and normalized as to that in WT MEFs. e OCRs of WT and DDHD2 KO MEFs were measured as described under Materials and methods. The scale bars for whole cell panels are 10 μm. Data represent means ± SEM for three (b, d, e) and five (c) independent experiments, respectively. *P < 0.05, **P < 0.01, Student’s t-test