Fig. 4
Areg−/− Treg cells induce less severe intestinal fibrosis. (A-F) CD45RBhi CD4+ T cell adoptive transfer model were used to induce chronic colitis in Rag−/− mice. Meanwhile, splenic CD4+ T cells were isolated from WT and Areg−/− mice and cultured under Treg condition for 5 days. On day 5, CD45RBhi CD4+ T with WT or Areg−/− Tregs were transferred into Rag−/− mice. The mice were killed 6 weeks after cell transfer, and the severity of intestinal inflammation and fibrosis were determined (n = 6/group). (A) Disease severity was measured by histopathology and pathological scores. Scale bars, 200 μm. (B) Il1b, Il6, and Tnf levels in in colonic tissues were measured by qRT-PCR. (C) Intestinal fibrosis was determined by Masson staining. (D) Col1a1, Col6a1 and Col6a3 levels in colonic tissues were measured by qRT-PCR. (E) α-SMA expression in colon tissues from WT Treg and Areg−/− Treg transfer mice was determined by western blot. (F) Colon tissues were stained with immunofluorescence. α-SMA layer thickness were analyzed. Representative data from 2 independent experiments with similar results. (A-F) Unpaired Student’s t-test. *P < 0.05; **P < 0.01; ***P < 0.001; ns: not significant