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Fig. 2 | Respiratory Research

Fig. 2

From: Combination of glycopyrronium and indacaterol inhibits carbachol-induced ERK5 signal in fibrotic processes

Fig. 2

Effect of GLY and/or IND on CCh- or TGF-β1-augmented gel contraction and α-SMA expression in HFL1 cells. Sub-confluent fibroblasts were placed in serum-free (SF)-DMEM for 24 h. Cells were pre-treated with GLY and IND for 30 min, casted into collagen gels, and released into SF-DMEM containing various concentrations of GLY and/or IND with or without CCh (a, b, e) or TGF-β1 (c, d, g). Gel size was measured on day 3. Vertical axis: gel size expressed as a percentage of control. Horizontal axis: conditions. All values represent the mean ± SEM of three separate experiments, each performed in triplicate. For α-SMA assay, cells were grown to sub-confluence in monolayer culture and serum deprived for 24 h. Cells were then stimulated with CCh or TGF-β1 in the presence or absence of GLY and/or IND. After 48 h of incubation with CCh and 24 h of incubation with TGF-β1, cells were harvested for immunoblotting using a α-SMA antibody (f, h). Vertical axis, relative intensity of α-SMA/β-actin ratio; Horizontal axis, conditions. Representative immunoblots are shown at the top of each panel. All values represent the mean ± SEM in three different strains. P < 0.05, † † P < 0.01, compared with solvent control. *P < 0.05, **P < 0.01, compared with stimulus

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