Inhibiting phosphorylation of Akt at Thr308 accompanied by decreased p‐eNOS Ser1177 expression. HUVECs were treated with GSK2334470 at different concentrations (0.01, 0.1, 0.5, 1 μm) or for different durations (2, 5, 10 min) to explore the effects of GSK2334470 on p‐eNOS Ser1177 expression. (A) Western blot and quantitative analysis of the protein expressions of p‐Akt Thr308, p‐Akt Ser473, and total‐Akt in HUVECs treated with different concentrations of GSK2334470 for 30 min. (B) Western blot and quantitative analysis of the protein expressions of p‐Akt Thr308, p‐Akt Ser473, and total‐Akt in HUVECs treated with GSK2334470 (1 μm) for different durations. (C) Western blot and quantitative analysis of the protein expressions of p‐eNOS Ser1177, p‐eNOS Thr495, and total‐eNOS in HUVECs treated with GSK2334470 (1 μm) for different durations. The relative levels of p‐Akt Thr308, p‐Akt Ser473, p‐eNOS Ser1177, and p‐eNOS Thr495 were quantified as the ratios of p‐Akt Thr308/Akt, p‐Akt Ser473/Akt, p‐eNOS Ser1177/eNOS, and p‐eNOS Thr495/eNOS, respectively. Data were expressed as mean ± SE, n = 4. Data were analyzed with one‐way ANOVA followed by Tukey's multiple comparisons test. *P < 0.05 vs. Control group; ***P < 0.001 vs. Control group.