Extended Data Fig. 6: Effects of PI3P and PI4P on mTORC1 activity and lysosomal localization of mTOR with Raptor membrane-interaction mutants.
From: Structural basis for mTORC1 activation on the lysosomal membrane
a, In vitro kinase assay showing mTORC1 kinase activity with different composition of negatively charged lipids. The quantification is shown for 3 repeats. b, iRaptor KO MEFs, untreated or treated with 0.5 μM 4-hydroxy-tamoxifen (4-OHT) for 48 h, were transfected with either WT, F1296E, or F1296/M1297E Raptor. 24 h after transfection, cells were starved of amino acids for 60 min, then re-stimulated with amino acids for 30 min. The cells were analyzed by immunofluorescence and quantified for mTOR-LAMP1 colocalization using Manders’ colocalization coefficient. Results are mean ± s.e.m.; n = 3 (One-way ANOVA, Dunnett’s multiple comparisons test; ns: not significant). Scale bar, 10 μm.
