Extended Data Fig. 3: Plasma membrane V-ATPase regulates cholesterol distribution and macropinocytosis.
From: Plasma membrane V-ATPase controls oncogenic RAS-induced macropinocytosis
a, Fluorescence micrographs of HeLa control and HeLa HRAS(G12V) or T7-KRAS(G12V) cells immunostained with V1A antibody. b, Validation of V1A antibody for immunofluorescence. Fluorescence micrographs of V1A immunostaining of HeLa HRAS(G12V) cells transfected with the indicated siRNAs. c, Fluorescence micrographs of V1A immunostaining and filipin labelling of HeLa KRAS(G12V) cells transfected with the indicated siRNAs. d, Quantification of TMR–dextran uptake in mutant RAS cell lines transfected with siV0a3 in the presence or absence of exogenous cholesterol. Images (a–c) are representative of three biological replicates. Scale bars, 10 μm. In b, c, dashed lines delineate the cell and/or nucleus. Data in a, c, are mean ± s.e.m. representing the fraction of cells that display V1A plasma membrane localization. At least 500 (a, c, d) cells were quantified in each biological replicate (n = 3); data are mean ± s.e.m.; unpaired two-tailed Student’s t-test.
