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

FIGURE 8.

FIGURE 8

Gene silencing of HIF-1α and/or HIF-1β does not suppress the IL-6/STAT3 signaling in cultured human myoblasts. Human myoblasts were treated with siRNA directed against HIF-1α (siHIF1α) and/or HIF-1β (siHIF1β). 44 h after application of siRNA cells were serum-starved for 4 h and then analyzed for (A) HIF-1α mRNA, (B) HIF-1β mRNA, and (C) IL-6 mRNA by the quantitative PCR or (D–G) treated with 50 ng/ml interleukin-6 (IL-6) for 15 min and analyzed for (D) HIF-1β, (E) the total STAT3 and (F) phospho-STAT3 (Tyr705) by immunoblotting. Graph in panel (G) shows the phospho-STAT3:STAT3 ratio. Two concentrations of scrambled siRNA (siSCR) were used as controls: 5 nM (siSCR1; control for siHIF-1α) or 45 nM (siSCR2; control for siHIF-1β and siHIF-1α + 1β). mRNA of target genes is expressed relative to mRNA of a reference gene (ACTB). Results are means with SEM (n = 3–4). *p < 0.05 vs. siSCR1; #p < 0.05 vs. siSCR2; §p < 0.05 vs. respective basal.