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Fig. 7 | Respiratory Research

Fig. 7

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

Fig. 7

Inhibition of ERK5 resulted in the suppression of CCh- or TGF-β1-stimulated gel contraction and α-SMA expression in lung fibroblasts. Sub-confluent HFL1 cells were pre-treated with ERK5 inhibitor (BIX02189), ERK1/2 inhibitor (PD98059), and Rho kinase inhibitor (Y27632) for 30 min followed by being casted into collagen gels and treated with CCh or TGF-β1. 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 in three separate experiments, each performed in triplicate (a). Sub-confluent HFL1 cells were serum deprived for 24 h followed by stimulation with CCh or TGF-β1 for 48 h. Cells were then harvested for α-SMA immunoblotting. Vertical axis, relative intensity of α-SMA/β-actin ratio; Horizontal axis, conditions. All values represent the mean ± SEM in the three different strains (b). Lung fibroblasts from non-smokers and subjects with COPD were grown to sub-confluence and pre-treated with BIX02189 for 30 min followed by being casted into collagen gels and released into SF-DMEM with CCh (c) or TGF-β1 (d) in the presence or absence of BIX02189. Gel size was measured on day 3. Vertical axis: gel size expressed as a percentage of control. Horizontal axis: conditions. Each pair of symbols connected by a line represented data from an individual cell strain and the mean of three separate experiments. Cells from non-smokers are indicated by open circles, cells from patients with COPD (GOLD I) by squares, and cells from patients with COPD (GOLD II) by closed circles. P < 0.05, †† P < 0.01, compared with solvent control. *P < 0.05, **P < 0.01, compared with stimulus. § P < 0.05, compared with non-smokers and COPD lung fibroblasts in the same stimulus group

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