GATOR1 is important for FLCN recruitment to lysosomes. (A) Western blots of WT and NPRL3 KO cells. Molecular masses are given in kilodaltons. (B) Spinning-disk confocal images of FLCN localization in WT and NPRL3 KO cells (HeLa) under basal and starved conditions. Bars, 10 µm. Insets are 7.4 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 = 3 experiments; 50–60 cells per condition; ****, P < 0.0001; ANOVA with Bonferroni’s post hoc test).