Autocrine Acetylcholine, Induced by IL-17A via NFκB and ERK1/2 Pathway Activation, Promotes MUC5AC and IL-8 Synthesis in Bronchial Epithelial Cells

Figure 8

Silencing of ChAT mRNA reduced Muc5AC expression and IL-8 release in 16-HBE cells stimulated with rhIL-17A 20 ng/mL for 24 h. Cells were stimulated and analyzed to evaluate (a) Muc5AC expression in unsilenced cells and in cells transfected with siRNA for ChAT or scrambled siRNA by flow cytometry. Bars represent mean ± SD of fluorescence mean intensity (FMI) of three separate experiments. (b) Representative flow cytometry of Muc5AC is shown. (c) IL-8 release (pg/mL, by ELISA) in unsilenced cells and in cells transfected with siRNA for ChAT or scrambled siRNA. The values shown are the mean ± SD for three separated experiments. (d) Knockdown efficiency (%) of ChAT mRNA on ChAT protein expression. Bars represent mean ± SD of three separate experiments. Statistical analysis was performed by ANOVA test followed by Fisher’s PLSD multiple comparison test.

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