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. 2018 Jun 19;32(12):6883–6897. doi: 10.1096/fj.201800413RR

Figure 7.

Figure 7

REDD1 expression attenuates JNK activity. A) HEK293E Tet-On HA-REDD1 cells were treated with Dox for the times indicated. Total REDD1, JNK, c-Jun, α-tubulin, GAPDH, and phosphorylation of JNK on Thr183/Tyr185 and c-Jun on Ser63 were assessed in whole-cell lysates by Western blot analysis. B, C) Quantification of phosphorylation of JNK (T183/Y185) (B) and c-Jun (Ser63) (C) at 0 and 6 h. D) REDD1-knockout (KO) MEFs were either dual transfected with p-TRE HA-REDD1 and pREV Tet-ON plasmids or transfected with an empty vector control plasmid (EV). Transfected MEFs were exposed to Dox for 24 h or the indicated time. E, F) Quantification of phosphorylation of JNK (T183/Y185) (E) and c-Jun (Ser63) (F) at 0 and 24 h. Protein loading was assessed by protein stain (Protein S). Blots shown are representative of results for 2 experiments; within each experiment, 2 independent samples were analyzed. Protein molecular mass (kDa) is indicated at the right of the blots. Results are expressed as means ± sem (n = 3). *P < 0.05, ***P < 0.01 vs. time 0 h.