Fig. 5.

RNF152 and USP4 regulate mTORC1 activation. a EGF-induced mTORC1 activation was measured in primary WT or RNF152^−/− MEF cells. RNF152 knockout was analyzed via genomic DNA PCR in Supplementary information, Fig. S3s. b Overexpressing RNF152, but not the E3 ligase dead mutant RNF152-CS, reduced EGF-induced mTORC1 activation in HEK293T cells. c The activation of mTORC1 induced by EGF was analyzed in Rheb-WT or Rheb-K8R knockin HEK293T cells. d, e Activation of mTORC1 induced by Rheb-K8R was resistant to RNF152 (d) and TSC2 (e). f Overexpressing USP4-WT, not USP4-CS, enhanced EGF-induced mTORC1 activation. g EGF-induced mTORC1 activation was detected in WT and USP4 deficient primary MEF cells. h, i mTORC1 activation was measured after treatment with different doses of Vialinin A for 4 h in HEK293T cells. j The effects of different USP4 mutants (USP4-WT, S445A, S445D) on mTPRC1 activation