Fig. 8
From: USP32 regulates late endosomal transport and recycling through deubiquitylation of Rab7
USP32 regulates extraction of membranes from Rab7-positive endosomes. a, b Bud/tubule resolution from Rab7 endosomes as a function of USP32. Select confocal frame zooms taken from time-lapses of live control (siCtrl) vs. USP32-depleted (siUSP32_2) MelJuSo cells stably expressing (a) GFP-Rab7 or (b) GFP-2KR (green) labeled with Lysotracker (magenta) are shown. Large arrows point to emerging buds (B) and tubules (T), small arrows point to nascent vesicles formed as a result of fission (F). Quantification: number (#) of resolved (white) and unresolved (black) buds and tubules observed, n = 2 independent experiments. Scale bars = 5 µm. See also Supplementary Movies 13–16. c Alterations in late endosome (LE) morphology in response to USP32 depletion as visualized by correlative light and electron microscopy (CLEM). GFP-Rab7-2KR (GFP-2KR) fluorescence (green) and transmission electron micrographs (TEMs) are shown; scale bars = 0.25 µm. d Comparison of GFP-Rab7-2KR-positive LE profile in siCtrl (black line) and siUSP32_2 (red line); x-axis: LE diameter in µm; y-axis: number of LE profiles. See also Supplementary Fig. 9
