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. 2016 Jan 11;291(12):6262–6271. doi: 10.1074/jbc.M115.672170

FIGURE 2.

FIGURE 2.

Specific mTOR activity signals collagen I expression in fibrotic mesenchymal cells with suppression of collagen I by AZD8055 but not rapamycin. A, Fib-MCs (n = 7) were treated with rapamycin (varying doses) for 24 h, with collagen I and phosphorylation of S6K1 (Thr-389), 4E-BP1 (Thr-37/46), and AKT (Ser-473) measured by Western blot analysis. B, Non-Fib MCs (n = 6) were treated with rapamycin (250 nm) for varying times (24–72 h) and 4E-BP1 (Thr-37/46), AKT (Ser-473), and collagen I expression measured by Western blot analysis. C, Fib-MCs (n = 7) were treated with AZD8055 for 24 h and phosphorylation of S6K1 (Thr-389), 4E-BP1 (Thr-37/46), and AKT (Ser-473) measured by Western blot analysis. D, effect of AZD8055 treatment on collagen I protein expression in Fib-MCs over varying time intervals of treatment is shown (n = 7). Densitometric analysis here is presented as percent of untreated control. Approximately 80% suppression in collagen I protein expression was induced by treatment with AZD8055, and this effect was persistent at 48 and 72 h. E, time course of collagen I suppression by AZD8055 in Fib-MCs (n = 8). F, acid-soluble collagen produced by Fib-MCs in in vitro tissue culture condition was quantitated by Sircol Collagen Assay in the presence of AZD8055 (n = 8). **, p < 0.01; ***, p < 0.001.