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. 2022 Mar 31;20(3):e3001594. doi: 10.1371/journal.pbio.3001594

Fig 5. RagC is necessary for AA-stimulated TFE3 phosphorylation and localization to lysosome.

Fig 5

(A) Control C2C12 cells and cells lacking RagC were maintained for 60 minutes in medium lacking AAs and then returned to complete media for the indicated times, followed by immunoblotting as indicated. Phospho-TFE3/totalTFE3 and phospho-S6K/totalS6K were quantified and normalized to the 15-minute time point of NTC (control). (B) Control C2C12 cells and cells lacking RagC were cultured with 250 nM Torin1 for 15 minutes, followed by immunohistochemistry for subcellular localization of TFE3 and LAMP2, a marker of the lysosome. (Scale bar: 10 μm). On the right: correlation by Pearson’s R of LAMP2 and TFE3 staining. (C) Quantitative PCR evaluation of expression of the indicated genes (normalized to the average expression of HPRT, TBP, and 36B4 as controls). **p < 0.01, ***p < 0.001, ****p < 0.0001 by Student t test. The data underlying all the graphs shown in the figure is included in S1 Data. AA, amino acid; KO, knockout.