Extended Data Figure 7: Lysosome-targeted WDR59 suppresses mTORC1 signalling in SZT2-deficient cells.

a–d, h, Control (a, b), SZT2-deficient (c, d), or SZT2- and WDR59-deficient (h) HEK293T cells were transfected with the indicated constructs and selected with puromycin. These cells were deprived of amino acids for 1 h and stimulated with amino acids for 20 min where indicated. Total cell lysates were analysed by immunoblotting. e, Control or WDR59-deficient HEK293T cells were analysed as in a. f, Primary Wdr59^+/+ or Wdr59^−/− MEFs were analysed as in a. g, mRNA levels of Wdr59 in Wdr59^+/+ and Wdr59^−/− MEFs were measured by qPCR and normalized to that of β-actin. AU, arbitrary units. HA, haemagglutinin. Data (a–h) are representative of three independent experiments.