Figure 4. NCOR1 is essential for Treg cell-mediated protection in adoptive CD4⁺ T cell transfer colitis.

(a) Experimental protocol for adoptive transfer colitis. Control groups received naïve CD4⁺ T cells only or no cells at all (not shown). (b) Weight scores in percentages of initial body weight during the course of colitis in Rag2^−/− recipient mice are shown. Data show the summary of at least eight mice (except control groups with six mice) of three independent experiments. *p<0.05, **p<0.01, and ***p<0.001 (two-way ANOVA analysis followed by Tukey’s multiple-comparisons test). For simplicity, significant differences are shown only between mice that received wild-type (WT) and NCOR1-cKO Treg cells. Of note, non-injected control mice gained significantly more weight (starting from around week 5) compared to all other groups. (c) Summary showing colon length from the various Rag2^−/− recipient mice. (d) Colon swiss rolls were processed for hematoxylin and eosin (H&E) staining. The pictures in the bottom represent a 5x magnification of the black rectangle framed section in the top pictures. Magnification: 40x and 200x. Scale bar = 100µm. One representative picture is shown per condition.

Figure 4—figure supplement 1. In vitro characterization of wild-type (WT) and NCOR1-cKO Treg cells.

(a) Flow cytometric analysis of splenocytes isolated from WT and NCOR1-cKO mice showing expression of TGFβ in CD4⁺FOXP3⁺ Treg cells after stimulation with PMA/ ionomycin for 4hr. (b) Summary showing TGFβ expression (gMFI) in FOXP3⁺ Treg cells of all mice analyzed as described in (a). (c) Histograms showing the proliferation of naïve CD4⁺ T cells co-cultured at different ratios with either WT or NCOR1-cKO Treg cells. (d) Summary showing the percentage of proliferating CD4⁺ T cells that have been cultured at different ratios with either WT or NCOR1-cKO Treg cells as described in (c). (c) Numbers indicate the percentages of cells in the respective gates. (b,d) Each symbol indicates one mouse or sample. (b) Horizontal bars indicate the mean. *p<0.05. Unpaired two-tailed Student’s t-test. Data are representative (a,c) or show a summary (b,d) of four (a) or at least two independent samples (c,d) that were analyzed in one experiment (a,b) or in two (c,d) independent experiments.