Extended Data Fig. 2: DMiro Protein Levels in MICOS Mutants.

(a) Lysates of 2^nd instar larvae (dMIC19^null), pupae (dMIC60^null), or 14-day-old adults (RNAi) were immunoblotted as indicated. The band intensity of each marker is normalized to that of ATP5β or β-actin from the same blot and graphed as relative change compared to control. Wild-type: w¹¹¹⁸. n=3 (Top panel: anti-DMiro for WT; and Middle panel) and 4 (the rest) independent experiments. Two-sided Mann-Whitney Test (Lower panel) and T Test with Welch’s correction (Top and Middle). n.s.: not significant. p=0.0009, 0.0005, 0.0096 for dMIC19 null, 0.0319, 0.0351, 0.0173 for dMIC60 null. The rest of the precise p values are in Source Data. (b) Mitochondrial (Mito) and cytosolic (Cyto) fractions from 20-day-old flies were immunoblotted as indicated. The DMiro band intensity is normalized to that of VDAC from the same blot. n=3. Two-sided T Test with Welch’s correction. p=0.0283, 0.0102. (c) Whole-body lysates of 20-day-old flies were immunoblotted as indicated. The band intensity is normalized to that of β-actin from the same blot and graphed as relative change compared to control (Actin-GAL4, gray bar) within the same experiment. n=5 flies per experiment, 4 independent experiments. p=0.0286. (d) The mRNA levels of DMiro were determined by RT-qPCR in 20-day-old flies, normalized to RP49 from the same experiment, and graphed as relative change compared to respective controls (white boxes) within the same experiment. n=4 independent experiments. p=0.0286. (c-d) Two-sided Mann-Whitney Test. (a, d) Boxes show 25th/75th percentiles, whiskers are the minimum and maximum, and middle line is median. (a, b, c) Data are presented as Mean±S.E.M. with dots.

Source data