1. MOSPD2 silencing affects interorganelle contact sites organization
MOSPD2 silencing affects interorganelle contact sites organization AWestern blot analysis of MOSPD2, VAP‐A, and VAP‐B proteins in control HeLa cells (WT) and in HeLa cells transfected with a control siRNA (siCtrl), and with siRNA targeting MOSPD2 (siMOSPD2), VAP‐A, and VAP‐B (siVAP‐A/VAP‐B), and MOSPD2, VAP‐A, and VAP‐B (siMOSPD2/VAP‐A/VAP‐B).B–F TEM images of control HeLa cells (B: HeLa; C: HeLa/siCtrl) and MOSPD2 (D: siMOSPD2), VAP‐A, and VAP‐B (E: siVAP‐A/VAP‐B), and MOSPD2‐, VAP‐A‐, and VAP‐B (F: siMOSPD2/VAP‐A/VAP‐B)‐silenced HeLa cells. An interpretation scheme representing contacts between organelles is shown on the right; the ER, endosomes, and ILV are in dark, light, and medium gray, respectively. Mitochondria and Golgi are in pink and light green, respectively. Scale bars: 500 μm.G–I Quantification by stereology of ER–mitochondria (G), ER–endosome (H), and endosome–endosome (I) contacts. The percentages of mitochondria perimeter in contact with the ER (G), endosome perimeter in contact with the ER (H), and endosome perimeter in contact with an endosome (I) are shown as means and error bars (SEM). (G) 200 (HeLa), 208 (siCtrl), 199 (siMOSPD2), 202 (siVAP‐A/VAP‐B), 199 (siMOSPD2/VAP‐A/VAP‐B) mitochondria from 8, 10, 8, 12, and 12 cells, respectively, were analyzed. (H, I) 202 (HeLa), 189 (siCtrl), 148 (siMOSPD2), 141 (siVAP‐A/VAP‐B), 221 (siMOSPD2/VAP‐A/VAP‐B) endosomes from 9, 19, 10, 14, and 12 cells, respectively, were analyzed. Kruskal–Wallis with Dunn's multiple comparison test (*P‐values < 0.05; ***P < 0.001; ****P < ). Source data are available online for this figure.
Thomas Di Mattia et al. EMBO Rep. 2018;19:e45453
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