Figure 6.

Adenosine inhibition of gap closure can be reversed by treatment with an adenosine receptor antagonist. (a) Exogenous adenosine inhibits esophageal keratinocyte wound healing. The effect of adenosine (1–100 µM) on the percentage of covered gap area in WT and TRPV4-KO keratinocyte cultures was examined. Cells were also treated with 10 nM MRS1754 (selective A_2B adenosine receptor antagonist) to examine its effect on in vitro wound healing inhibition mediated by 1 µM adenosine. Data are presented as means ± SEM (n = 6–12). *p < 0.05; **p < 0.01 compared with WT, ^‡p < 0.05; ^‡‡p < 0.01 compared with WT control, ^§§p < 0.01 compared with TRPV4-KO control, ^##p < 0.01 compared with WT 1 μM adenosine, ^¥¥p < 0.01 compared with TRPV4-KO 1 μM adenosine; as determined by one-way ANOVA followed by Tukey post-hoc test. (b) Adenosine receptor mRNA transcription was examined with ( +) and without (–) RT reaction. All adenosine receptor subtypes were transcribed in the esophageal mucosa of WT and TRPV4-KO mice, but A_2B adenosine receptor had apparently higher transcription compared to A₁, A_2A and A₃ adenosine receptors. Full-length gels are shown in Supplementary Fig. S7.