SETD2 mediates ROS reaction to control cell apoptosis and colon inflammation.
(A) Heat map of RNA-seq data to compare the gene expression in the IECs from Setd2Vil-KO and Setd2f/f mice treated for 4 d with DSS. Go term analysis of gene expression changes in the right.
(B) 8-OHdG staining from DSS-treated (5d) Setd2Vil-KO and Setd2f/f mice. Scale Bars: 50 μm.
(C) Histograms and MFI quantifications of ROS in IECs isolated from DSS-treated (5d) Setd2Vil-KO and Setd2f/f mice (n = 4 per genotype).
(D–H) 8-week-old Setd2Vil-KO and Setd2f/f mice were pretreated with NAC (100 mg/kg) every day for 4 days induced by intraperitoneal injection, and treatment was continued during DSS administration for 5 days.
(D) The mice were euthanized on days 10 to measure colon length (n = 6 per genotype). Scale Bars: 1 cm.
(E) H&E stained sections of colon tissues collected on day 10 from DSS-treated mice as indicated, and semi-quantitative scoring of the histopathology is shown in the right (n = 6 per genotype). Scale Bars: 100 μm.
(F) Relative mRNA expression levels of inflammatory mediators in whole colon from DSS-treated (5 d) mice as indicated (n = 5 per genotype).
(G) Cleaved caspase-3 staining of colon sections from DSS-treated (5 d) Setd2Vil-KO and Setd2f/f mice with or without NAC treatment (n = 5 per genotype). Scale Bars: 50 μm.
(H) Organoids were isolated from 8-week-old Setd2Vil-KO and Setd2f/f mice with or without NAC treatment, and cleaved caspase-3 staining (n = 4 per genotype). Scale Bars: 20 μm.
The data represent the mean ± S.E.M, and statistical significance was determined by a two-tailed Student's t-test. *p < 0.05, **p < 0.01, ***p < 0.001. N.S., Not Significant. MFI, Median Fluorescence Intensity.