Fig. 1: ITF2 expression is attributable to p65.

A Schematic diagram of DSS treatment to induce acute colitis in the mouse intestine. Western blot analysis of ITF2 in the colonic epithelial cells of WT mice treated with 3.5% DSS for 0–7 days. Mouse icon was created with BioRender (BioRender.com). n = 2, biologically independent experiment. B Phase contrast image (left panel) and western blot analysis of ITF2 in the colonic organoids treated with TNF (20 ng/ml) in a concentration-dependent manner (right panel, n = 2, biologically independent experiment). C Colo320DM cells were pretreated with the indicated inhibitors for 1 h, followed by TNF stimulation for 8 h. MG132 was treated without TNF. Specified protein expressions were traced. n = 3, biologically independent experiment. D Colo320DM cells were pre-incubated with indicated NF-κB inhibitors for 1 h and cultured with TNF for 8 h. The indicated proteins were traced and presented as immunoblots. Each data are expressed as normalized band intensity adjusted to α-tubulin, which serves as a loading control. The protein intensities were quantified by ImageJ software. n = 3, biologically independent experiment. E Colo320DM cells were overexpressed with p65 and p50, representative NF-κB subunits, in a concentration-dependent manner with or without ITF, and then ITF2 expression was evaluated. n = 2, biologically independent experiment. F Colon cancer cells were transfected with p65 and ITF2 expression was compared. n = 3, biologically independent experiment. G Representative immuno-stained images of ITF2 and p65 expression in acute colitis-induced colon tissues. The data are representative of three independent experiments. H Epithelial expression levels of ITF2 were analyzed and compared (n = 5, 10, 10 respectively, biologically independent animals., **P = 0.0027, **P = 0.0007, *P = 0.0122). I Epithelial expression levels of p65 were analyzed and compared (n = 5, 10, 10 respectively, biologically independent animals., **P = 0.0007, **P = 0.0006, **P = 0.0006). J Linear regression between ITF2 and p65 in each group of mice (Spearman r = 0.8506, **P < 0.001). K Stable cell lines were established with small hairpin RNA targeting ITF2 and treated with or without TNF. The indicated proteins were traced and presented as immunoblots. n = 2, biologically independent experiment. Scale bars, 100 μm. In all immunoblot analyses, the data are representative of three independent experiments. All results are presented as means ± SEM. Statistical significance was determined by the two-tailed Mann–Whitney U test (H, I) and the two-tailed Spearman correlation test (J). Source data are provided in the Source Data file.