Figure 4.

GW501516 and GSK0660, when administered alone or in combination, suppressed the TGF-β₁-induced upregulation of Cx43 in HBF populations. HBFs derived from asthmatic patients were cultured in a serum-free medium containing GW501516 and GSK0660 alone or in combination, and in the absence or presence of TGF-β₁ (5 ng/mL) for four days. (A) Representative images of HBFs immunostained for Cx43 (red) with DNA visualization (blue) are presented. Scale bar = 50 µm. Cx43 levels in HBFs were determined via (B) the quantification of the fluorescence signal in the collected images (n = 5) or via (C) Western blot analyses (n = 1, densitometry is presented above the bands). (D) The relative expression of GJA1 (the Cx43-encoding gene) in HBFs (n = 3) cultured for 24 h under the conditions described above was determined using real-time PCR. The results are presented as 2^−ΔΔCt mean values in relation to the control gene (GAPDH). All results are presented as the mean ± SEM. Statistical significance was tested using the nonparametric Kruskal–Wallis test with Dunn’s multiple comparisons post hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001.