Compartmentalization of mitochondria is independent of the fission machinery. (A–D) Dual-color FLIP in fis1∆ cells expressing GFP-tagged mitochondrial proteins and matrix-mCherry. Representative images, pooled quantification data of GFP FLIP, and t_X of GFP FLIP from three independent experiments are shown. Photobleach was applied in the GFP and mCherry channels as indicated with white circles. (Note here that the bleaching and imaging conditions were changed and the graphs cannot be directly compared with Fig. 2). (A)fis1∆ cells expressing Tom20-GFP and matrix-mCherry (n = 32 cells, n ≥ 10 cells for each experiment). (B)dnm1∆ cells expressing Tom20-GFP and matrix-mCherry (n = 28 cells, n ≥ 7 cells for each experiment). (C)fis1∆ cells expressing Hem1-GFP and matrix-mCherry (n = 21 cells, n = 7 cells for each experiment). (D)fis1∆ cells expressing Atp1-GFP and matrix-mCherry (n = 25 cells, n ≥ 8 cells for each experiment). Images are a sum projection of five z-stacks taken at 0.5 μm intervals. Scale bar: 3 μm. Data from three independent clones were pooled to obtain the bleaching curves. Shadows represent mean ± SD. Error bar: mean ± SE. Welch’s two-tailed t test was applied to compare the t_X in the mother and bud.