Fig. 2

NUAK1 inhibition induces mTOR accumulation at the lysosome. A Representative confocal images of mTOR under NUAK1 inhibition. MDA-MB-231 cells were serum-starved overnight followed by 90 min of pretreatment with DMSO or HTH-01-015 (10 µM) before EGF stimulation for 0, 10, and 30 min. Green, mTOR; Blue, nuclei. B Representative confocal images of mTOR under NUAK1 inhibition. MDA-MB-231 cells were serum-starved overnight followed by 90 min of pretreatment with DMSO or HTH-01-015 (10 µM) and EGF-stimulated for 60 min. Green, mTOR; Blue, nuclei. C Quantification of the number of dots (upper) and intensity (lower) from B. Each bar represents the mean ± SD, Student t test. D Representative confocal images of mTOR and Lamp1-RFP after NUAK1 inhibition from non-stimulated or EGF-stimulated MDA-MB-231 cells for 60 min. Left, merge; Right, mTOR and Lamp1-RFP images. Green, mTOR; Red, Lamp1-RFP; Blue, nuclei. E Representative confocal images of endogenous mTOR (N-19) and Lamp1 in MDA-MB-231 cells after NUAK1 inhibition from non-stimulated or EGF-stimulated cells for 60 min. Left, merge; Right, zoom in. Green, mTOR (N-19); Red, Lamp1; Blue, nuclei. F Quantification of mTOR/Lamp1 co-localization from E. Each bar represents the mean ± SD, Student t test