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. 2020 Sep 25;11:566584. doi: 10.3389/fphys.2020.566584

FIGURE 3.

FIGURE 3

Ouabain stimulates the phosphorylation of Akt and p70S6K in cultured human myotubes. (A–D) Human myotubes were treated with or without 50 nM ouabain (OUA) in Advanced MEM with 2% FBS for 20 h and in serum-free Advanced MEM for 4 h. Cells were treated with tocilizumab (100 μg/ml, TCB) during the last 4 h and with interleukin-6 (50 ng/ml, IL-6) during the last 15 min. (E–L) Human myotubes were treated in serum-free Advanced MEM with different concentrations of OUA for 24 h (E–H) or with 50 nM OUA for 1–24 h (I–L). (A,E,I) The total Akt, (B,F,J) phospho-Akt (Ser473), and (D,H,L) phospho-p70S6K (Thr389) were measured by immunoblotting. Graphs in panels (C,G,K) show the phospho-Akt:Akt ratio. Results are means with SEM (n = 4–8). *p < 0.05 vs. Basal, ¥p = 0.058.