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. 2018 Dec 14;38(3):e99548. doi: 10.15252/embj.201899548

Figure 4. Phosphorylation of RagC alters mTORC1 stability but not the localization of mTOR .

Figure 4

  1. Empty vector (EV), HA‐RagC WT, 3A, or 3E was transiently expressed in HEK‐293E cells. RagC and lysosome maker, Lamp1, were monitored by confocal fluorescence microscopy. Scale bar, 10 μm.
  2. Empty vector (EV), HA‐RagC WT, or 3A was transiently expressed with scramble or RagC and RagD shRNA in HEK‐293E cells. mTOR and HA‐RagC were monitored by confocal fluorescence microscopy. Scale bar, 10 μm.
  3. Empty vector (EV), Flag‐RagC WT, 3A, or 3E was transiently expressed with HA‐RagA in HEK‐293E cells. Total cell lysates and amounts of mTOR, raptor, and HA‐RagA in the Flag‐RagC immunoprecipitates were analyzed by Western blot. Graphs show mean ± SEM of quantitative analyses of Western blots (*P < 0.05, two‐tailed Student's t‐test, n ≥ 3).
  4. Expression of HA‐RagC WT, 3A, or 3E was induced by overnight addition of tetracycline in HeLa cells. Total cell lysates (TCL) and amount of mTOR in the raptor immunoprecipitates were analyzed by Western blot. Graphs show mean ± SEM of quantitative analyses of Western blots (***P < 0.001, Student's t‐test, n = 3).

Source data are available online for this figure.