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

Figure 7

Effect of Hemicholinium-3 (HCh-3) or Tiotropium on Muc5AC production in 16-HBE cells. Cells were incubated for 1 h with HCh-3 (50 μM) or Tiotropium (100 nM) before addition of rhIL-17A 20 ng/mL for 24 h to evaluate ((a)–(d)) Muc5AC protein expression by flow cytometry. Bars represent mean ± SD of fluorescence mean intensity (FMI) of three separate experiments. Data were plotted as fold change compared to untreated cells. Representative flow cytometry of Muc5AC is shown. ((e)-(f)) Muc5AC protein expression by western blot. Bars represent mean ± SD of arbitrary densitometric units (ADU) of three different experiments. Representative western blot analysis of Muc5AC protein is shown. Statistical analysis was performed by ANOVA followed by Fisher’s PLSD multiple comparison test. The black curve represents the anti-IgG isotype negative control antibody.

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