Figure 2.

Effect of proinflammatory mediators on Cx43 expression in CFs. (A) Quiesced cells were stimulated with 10% FBS, Ang II (500nM), TGFβ1, IL-1β, IL-1α, TNFα (all 10ng/mL) or IL-6 in the presence or absence of the sIL-6R (both 50 ng/mL) for 24 h. Cells maintained in 0.5% FBS media were used as a control. Data represented as relative quantification (RQ) ± rq_max normalised to UBC (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 vs. control (one-way repeated measures ANOVA with Dunnett's multiple comparison test). (B) Immunostaining of Cx43 (green) in CFs maintained in 0.5% FBS media (control) or treated with 10ng/mL IL-1β (n=3). Nuclei were counterstained with DAPI (blue). Images were normalised to an IgG control shown in the upper right panel. The areas surrounded by a red box are expanded within the merged image or in the lower right panel for control and IL-1β stimulated cells respectively. White arrows indicate punctate Cx43 expression (green: rabbit polyclonal anti-Cx43). Images were taken at x25 magnification. Scale bar represents 100 μm. (C) Quantification of immunofluorescence normalised to the total cell number present in the images. At least 2 images per condition were analysed in each of the 3 independent experiments. **P < 0.01 vs. control (unpaired, two tailed t-test).