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

a, b, Control (sgGFP) or 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 where indicated. Total cell lysates were analysed by immunoblotting. c, The size of control and SZT2-deficient HEK293E cells was determined by flow cytometry. d, HEK293T cells stably overexpressing Flag–RAP2A or Flag–SZT2 cells were analysed 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 where indicated. The localization of mTOR and LAMP2 was determined by immunostaining. f, Quantification of the co-localization between mTOR and LAMP2. Data represent mean ± s.d. 100 cells were analysed from each condition (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 where indicated. Total cell lysates were analysed by immunoblotting. Data (a–e, g) are representative of three independent experiments.