Extended Data Figure 1. SZT2 is essential for mTORC1 inactivation upon nutrient deprivation.

a, b, Control, SZT2-deficient HeLa cells (a) or HEK293T (b) cells were deprived of amino acids and/or glucose for 1 h and stimulated with amino acids and/or glucose for 10 min when indicated. Total cell lysates were analyzed by immunoblotting. c, The size of control and SZT2-deficient HEK293E cells were determined by flow cytometry. d, HEK293T cells stably overexpressing FLAG-Rap2A or FLAG-SZT2 cells were analyzed as in (a). e, Control or SZT2-deficient HeLa cells were deprived of amino acids for 1 h and stimulated with amino acids for 10 min when indicated. The localization of mTOR and Lamp2 was determined by immunostaining. f, Quantification of the colocalization between mTOR and Lamp2. Data represent mean ± SD. 100 cells were analyzed from each condition with two-tailed unpaired t-test. ns, not significant. g, Control or SZT2-deficient HEK293T cells were transfected with the indicated constructs and selected with puromycin. Cells were deprived of amino acids for 1 h, and stimulated with amino acids for 10 min when indicated. Total cell lysates were analyzed by immunoblotting. Data (a – e, g) are representatives of three independent experiments.