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. Author manuscript; available in PMC: 2023 Jan 13.
Published in final edited form as: Nature. 2022 Jul 13;607(7919):610–616. doi: 10.1038/s41586-022-04939-z

Extended Data Figure 1: GATOR2 integrity and composition are insensitive to amino acid availability.

Extended Data Figure 1:

(a) Amino acids do not regulate the size of the GATOR2 complex. Elution fractions from size-exclusion chromatography of transfected GATOR2 or endogenously-tagged GATOR2 were analyzed by SDS-PAGE. Gels were stained with Coomassie Blue or SYPRO Ruby, as indicated. n.s. indicates a non-specific co-purifying contaminant. Lane labels indicate elution volumes from TSKgel G4000SWxl column.

(b) Analyses of endogenously tagged-WDR59 and -DEPDC5 expression in cell lysates.

(c) Analysis of mTORC1 signaling in cell lysates from which endogenously-tagged GATOR2 was purified in (a).

(d) Amino acids do not regulate the subunit composition or stoichiometry of endogenous GATOR2. Peak elution fractions from size-exclusion chromatography of endogenously-tagged GATOR2 were analyzed by SDS-PAGE and the gel was stained with SYPRO Ruby.

(e) The subunit composition of GATOR2 and the interaction between GATOR2 and GATOR1 or KICSTOR are not regulated by amino acids. Anti-FLAG immunoprecipitates were prepared from HEK293T cells with the indicated genotype and were analyzed by immunoblotting for the indicated proteins.

Data in (a)-(e) are representative of two independent experiments. For gel source data, see Supplementary Figure 1.