Figure 1. IL-10 is synthesized and released as a response to intestinal mucosal injury.

(A) Scratch wound-healing assay using IEC monolayers. rhIL-10 was added to wounded IECs, and wound widths were determined 0, 12, and 24 hours after injury (**P < 0.01 and ***P < 0.001, n = 5, mean ± SEM). (B) IEC expression of IL-10Rα was analyzed by qPCR and Western blotting. (C) Scratch wound-healing assay in IEC monolayers. Cells were transfected or not transfected with either a scramble siRNA or IL-10Rα siRNA, and wound widths were determined 0 and 24 hours after wounding (***P < 0.001, n = 5, mean ± SEM). Colonoscopy-based biopsy wounds (2-mm punch biopsies) were generated in C57BL/6 mice and collected on days 1–3 after injury. Intact tissue was used as a control. These samples were analyzed by qPCR for IL-10 kinetics in intestinal mucosal wounds (D), ELISA (E), and Western blotting (F). (D) Il10 qPCR of intact and wounded tissue on different post-injury days (*P < 0.05 and ***P < 0.001; n = 3, mean ± SEM). (E) Punch biopsy samples (2-mm) of resealing colonic wounds on different post-injury days and intact tissue were incubated overnight in complete DMEM. Supernatants were collected, and IL-10 secretion was analyzed by ELISA. (***P < 0.001; n = 3, mean ± SEM). (F) Lysates from wounded tissue on different post-injury days and intact tissue were immunoblotted for IL-10 (representative blot is shown, n = 3). Statistical comparisons were performed using ANOVA with Tukey’s multiple comparisons post test and a 2-tailed Student’s t test. IT, intact tissue; NT, nontransfected; Scr, scramble.