1. STARD3‐mediated cholesterol accumulation in endosomes does not alter cholesterol homeostasis
STARD3‐mediated cholesterol accumulation in endosomes does not alter cholesterol homeostasis Total free cholesterol quantification after total lipid extraction in control cells and in cells expressing STARD3 or the lipid binding mutant form of STARD3 (STARD3 MR/ND) (n = 3). Please note that control HeLa cells treated with MβCD have a significant depletion of total cholesterol (n = 2). Mean ± SD; ANOVA with Tukey's multiple comparison test.Western blot analysis of SREBP‐2 activation in controls (HeLa, HeLa/empty vector) or STARD3‐overexpressing HeLa cells. Cells were incubated for 2 h in DMEM culture medium supplemented with: 5% FCS (NT); 5% LPDS, 10 μM mevinolin (LPDS); 5% LPDS, 10 μM mevinolin, 1 mM MβCD (MβCD); 5% LPDS, 10 μM mevinolin, 500 μM cholesterol complexed to MβCD (MβCD‐Chol). The proteasome inhibitor MG132 (10 μM) was present in all conditions. P = precursor form of SREBP‐2; C = cleaved form of SREBP‐2. Lower panel: WB quantification in which cleaved SREBP‐2 is expressed as a fraction of total SREBP2 (P+C). Mean ± SD; n = 4 independent experiments; **P < 0.01, ***P < 0.001, ANOVA with Tukey's multiple comparison test. Source data are available online for this figure.
Léa P Wilhelm et al. EMBO J. 2017;36:
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