Extended Data Fig. 1: Mitochondrial integrity and pre-clearing efficiency.
From: MIROs and DRP1 drive mitochondrial-derived vesicle biogenesis and promote quality control
a) Representative confocal images of TOMM20+-MDVs in COS-7 cells. MDVs are positive for TOMM20 and negative for PDH. Scale bar = 5 µm. b) Representative confocal images of TOMM20+-MDVs in COS-7 cells expressing human TOMM20-3xHA and MAPL-h319L-Halo. Scale bar = 5 µm. The close-up demonstrates the unchanged cargo specificity of TOMM20+-MDVs in these cells. Orange circles = MDVs only positive for TOMM20. Scale bar = 1 µm. c) SDS-PAGE analysis of mitochondrial integrity after homogenization of cells by protease protection. Mitochondria were either untreated, swollen, or swollen and solubilized by Triton X-100 (TX100) in the presence of Proteinase K. OMM = outer mitochondrial membrane, IMS = intermembrane space. d) Quantification of the intermembrane space protein YME1L after Proteinase K treatment as a readout for the integrity of the outer mitochondrial membrane. Three experiments with single values (circles), averages (bars) and standard deviations (error bars) are shown (n = 3 biologically independent experiments). e-g) SDS-PAGE analysis (e) and quantifications (f, g) of pre-clearing efficiency through Halo link resin. Three experiments with single values (circles), averages (bars) and standard deviations (error bars) are shown (two-tailed unpaired Student’s t-test: ** = p ≤ 0.01, *** = p ≤ 0.001, n = 3 biologically independent experiments). Unprocessed blots and numerical source data including exact p values and test statistics are available in source data.
