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. 2017 Mar 10;8(19):31065–31078. doi: 10.18632/oncotarget.16075

Figure 3. SC-79 induced cytosolic activation of Akt antagonizes the inhibitory effect of ethanol on Ser473 in BMSCs.

Figure 3

(A) p-Akt-Ser473 levels in BMSCs decreased upon ethanol treatment while p-Akt-Thr308 status remained the same. SC-79 rescued p-Akt-Ser473 and p-GSK3β levels, impaired by ethanol or Ly294002 treatment. The western blot shows p-Akt, total-Akt, p-GSK3β, total-GSK3β, and β-actin as internal reference. (B) Western blot indicated that SC-79 could rescue β-catenin, impaired by Ly294002 treatment. β-actin served as internal reference. (C) SC-79 rescued the anti-osteogenic effect induced by Ly294002. The western blot shows COL1 and β-actin served as an internal reference. (D) Alizarin red staining was performed after BMSCs treated with control, Ly294002 (50 μM), SC-79 (10 μM) or their combination for 14 days. (E) Ethanol, Ly294002, SC-79 and their combinations blocked the translocation of Akt from cytoplasm to plasma membrane induced by IGF-1; however, the SC-79 independent cytosolic activation of Akt due to blockage effect by ethanol was not reversed. (F) Ly294002 treatment did not suppress the survival and proliferation of BMSCs. Values are shown as mean ± SD (n = 3).