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. Author manuscript; available in PMC: 2015 Jan 22.
Published in final edited form as: Sci Signal. 2014 Jul 22;7(335):ra68. doi: 10.1126/scisignal.2005103

Figure 3. REDD1 expression differentially alters the phosphorylation of Akt on Thr308 and Ser473.

Figure 3

(A-D, F-I) REDD1+/+ and REDD1−/− MEFs were serum starved and subsequently treated with the indicated concentrations of insulin. Relative phosphorylation of various proteins in the AktmTORC1 signaling pathway was assessed in whole cell lysates by Western blot analysis. Results represent mean phosphorylation relative to protein abundance ± SEM for (B) to (D) and (F) to (I). Dose response curves for REDD1−/− MEFs were significantly different (p<0.01) than REDD1+/+ MEFs for (B) to (D), (F), (G), and (I). (E) HEK293 Tet-On control cells (Control) and HEK293 Tet-On HA-REDD1 cells (HA-REDD1) were treated with doxycycline (Dox) as indicated. Phosphorylation of Akt on Thr308 and Ser473, TSC2 on Thr1462, PDK1 on Ser241, S6K1 on Ser229, as well as the abundance of each respective protein, HA-REDD1, and GAPDH was examined by Western blot analysis. (J) The relative phosphorylation of Akt on Thr308 and Ser473 in HEK293 Tet-On HA-REDD1 cells following treatment with doxycycline (Dox) as indicated. Blots in panel (A) and (E) are representative of results for three experiments (N=3); within an experiment, two independent samples were analyzed. Results represented as means ± SEM are shown in (J). *p<0.05 compared to 1 h.