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. Author manuscript; available in PMC: 2017 Aug 24.
Published in final edited form as: Nature. 2017 Feb 15;543(7645):433–437. doi: 10.1038/nature21378

Figure 4. Lysosome-targeted WDR59 or Sestrin2 inhibits mTORC1 signaling in the absence of SZT2 or GATOR1.

Figure 4

a, Cells, transfected with the indicated constructs and selected with puromycin, 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. b, Cells were deprived of amino acids for 1 h and stimulated with amino acids for 10 min when indicated, and analyzed as in (a). c, SZT2/WDR59/WDR24/MIOS-deficient HEK293T cells were transfected with lyso-FLAG-WDR59/Sestrin2. The localization of lyso-FLAG-WDR59/Sestrin2 was determined by immunostaining. d, e, Cells were transfected with the indicated constructs and selected with puromycin, and analyzed as in (a). f and g, Cells stably expressing the indicated proteins were analyzed as in (b). Data (a – g) are representatives of two independent experiments.