RagA and RagB are essential for starvation-dependent recruitment of FLCN to lysosomes. (A) Western blot analysis of WT and RagA/B KO HeLa cells. Molecular masses are given in kilodaltons. (B) Representative immunofluorescence experiment showing the effect of starvation on FLCN subcellular localization in WT and RagA/B KO HeLa cells. Bars, 10 µm. Insets are 12 µm wide. (C) Quantification of FLCN lysosome localization (percentage of cells with five or more FLCN spots that are also LAMP1 positive; mean ± SEM; n = 4 experiments, 76–80 cells per condition; ****, P < 0.0001; ANOVA with Bonferroni’s post hoc test).