Extended Data Fig. 6: PKA activation rescues RAS-induced macropinocytosis from sAC inhibition.
From: Plasma membrane V-ATPase controls oncogenic RAS-induced macropinocytosis
a, b, Effect of sAC inhibition (KH7) and rescue (KH7 + CA-PKA) on macropinocytosis, V-ATPase localization and cholesterol distribution in HeLa HRAS(G12V) cells. a, Quantification of TMR–dextran uptake following indicated treatments. b, Fluorescence micrographs of V1A immunostaining and filipin labelling following indicated treatments. c, d, Effect of sAC inhibition (KH7), plasma membrane V-ATPase inhibition (siV0a3), and rescue (KH7 + CA-PKA + siV0a3) on macropinocytosis, V-ATPase localization and cholesterol distribution in HeLa HRAS(G12V) cells. c, Quantification of TMR–dextran uptake following indicated treatments. d, Fluorescence micrographs of V1A immunostaining and filipin labelling following indicated treatments. e, Effect of KRAS inhibition (siKRAS) and rescue (siKRAS + CA-PKA) on macropinocytosis by quantification of TMR–dextran uptake in mutant RAS cell lines. Images in b, d are representative of three biological replicates. In b, d, dashed lines delineate the cell and nucleus; data are mean ± s.e.m. representing the fraction of cells that display V1A plasma membrane localization. Scale bars, 10 μm. At least 500 cells (a, c, e) were quantified in each biological replicate (n= 3); data are mean ± s.e.m.; unpaired two-tailed Student’s t-test.
