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. 2016 Aug 29;17(10):1471–1484. doi: 10.15252/embr.201642194

Figure 3. NFR‐mediated eEF2 phosphorylation is AMPK and mTOR independent.

Figure 3

  1. Schematic representation of signaling pathways targeted by the inhibitors used in this study.
  2. eEF2K WT and KO MEFs treated for 6 h with the indicated concentrations of NFR, rapamycin (Rapa), AICAR, or starved for the indicated time in PBS (Starv.), were analyzed by immunoblot using the indicated antibodies. Tubulin is used as a loading control.
  3. Potent mTOR inhibition does not impair NFR‐mediated eEF2 phosphorylation. WT MEFs were treated with the indicated concentrations of rapamycin or with vehicle (Mock). After 30 min, indicated doses of NFR were added and cells were incubated for an additional 6 h and analyzed for phosphorylated S6R and eEF2. Tubulin is used as a loading control.
  4. NFR‐mediated eEF2 phosphorylation is not affected in AMPKα1α2 dKO. AMPKα1α2 WT and dKO MEFs treated for 6 h with indicated concentration of NFR, rapamycin (Rapa), AICAR, or starved for 1 h in PBS (Starv.), were analyzed by immunoblot with the indicated antibodies. Anti‐total AMPKα antibody gives an unspecific band (*) with a slightly higher molecular weight in dKO cells. Tubulin is used as a loading control.
Data information: Each panel is representative of at least three independent experiments.Source data are available online for this figure.