MicroRNA-101-3p Suppresses mTOR and Causes Mitochondrial Fragmentation and Cell Degeneration in COPD

Figure 6

The effect of TNF-α on mTOR signaling pathway. (a) Western blots of TNF-α-induced short-term p70S6K phosphorylation (120 min.) in healthy nonsmokers (n = 3) and subsequent image analysis. Bars show the ratio of phosphorylated (p-) to total (t)-p70S6K as percentage of time point 0. (b) Western blots of TNF-α-stimulated long-term (48 hours) phosphorylation of (p)-p70S6K in ASMC of healthy nonsmokers (control, n = 3). (c) Western blots of TNF-α-induced long-term p70S6K phosphorylation in COPD-ASMC (n = 3) over 48 hours. (d) Image analysis of Western blots is presented in Figures 6(b) and 6(c). Bars compare the ratio of phosphorylation (p)-p70S6K to total (t-) p70S6K. (e) Image analysis of Western blots is presented in Figures 6(b) and 6(c). Bars show the ratio of t70S6K to GAPDH as percentage of time point 0. (f and g) Representative Western blots of TNF-α-induced p70S6K phosphorylation in ASMC of healthy nonsmokers (control) in the presence of different signaling pathway inhibitors (Rapamycin, PD98059, LY294002, and SB203580) at 24 hours. Similar results were obtained in three additional experiments. All values were calculated by comparison to untreated cells.

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