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. 2016 Jan;186(1):134–144. doi: 10.1016/j.ajpath.2015.09.013

Figure 6.

Figure 6

Stimulation by CRHR2 promotes colonic epithelial cell migration. A: Human CRHR2-transfected human colonic mucosal epithelial NCM460 cells (CRHR2) and EV transfected cells were stimulated with varying concentrations of Ucn2 with or without pretreatment of 1 μmol/L Ast2B, and migration was assessed with the slope (1/h) of data collected with xCELLigence software. B: Representative micrographs from CRHR2-overexpressing (CRHR2+) or EV cells treated with IL-6, Ucn2, or the combination after scratch assay at 0 and 48 hours. C: Pixel-based quantification of total wound area after scratch assay. D: Ucn2 and IL-6–induced STAT3 phosphorylation (Tyr705) in EV and CRHR2-overexpressing NCM460 cells, as assessed by Western blot analysis. Data are expressed as means ± SEM. n = 3 wells per group. ∗∗P < 0.01, ∗∗∗P < 0.001 versus control-treated cells on day 0; ††P < 0.01 versus Ucn2 treatment alone; †††P < 0.001 versus Ucn2-treated EV cells; ‡‡P < 0.01 versus Ucn2-treated CRHR2 cells; ‡‡‡P < 0.001 versus Ucn2-treated CRHR2 cells. Ast2B, astressin 2B; CON, control; CRHR2, corticotropin-releasing hormone receptor 2; EV, empty vector; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; M, molar concentration; pSTAT3, phosphorylated STAT3; STAT3, signal transducer and activator of transcription 3; Ucn2, urocortin 2.