Knocking down or ablating eEF2K elevates HSP90 protein levels.
A, lysates of A549 cells treated ±IPTG (to induce shRNA expression to knock down eEF2K) or eEF2K+/+ and eEF2K−/− MEFs were subjected to SDS-PAGE followed by WB analysis of the indicated proteins. B and C, quantification of HSP90 levels in A549 cells (B) or MEFs (C) from A. D, eEF2K+/+ and eEF2K−/− MEFs were cultured in low-glucose (5.5 mm) DMEM ± 10 mm 2-DG for 24 h before immunoblot analysis. E, quantification of HSP90 levels from D. P-eEF2, phosphorylated eEF2. *, 0.01 ≤ p < 0.05; **, 0.001 ≤ p < 0.01. Error bars represent S.D.