Fig. 7: Pharmacological inhibition of P2Y₁₄R ameliorates in DSS-induced colitis.

a Experimental flow chart. b Body weight and disease activity index evaluation of mice change during the disease process (n = 6 mice per group). c The length of colons from mice 7 days after DSS treatment (n = 6 mice per group). d The H&E staining in the colon tissues of DSS-treated mice (Scale bar = 200 μm). e Effects of P2Y₁₄R antagonists on mucosal barrier function as measured by serum level of FITC-dextran based on intestinal permeability method (n = 6 mice per group). f The immunofluorescent images of colon tissues stained with Claudin-1, Occludin and ZO-1, the principal components of tight junction (Scale bar = 200 μm). g The TEM images showed the typical characteristics of necroptosis in the colon tissues of DSS-treated mice. h Phosphorylation MLKL, PKA, CREB as well as its protein levels were analyzed by immunoblotting in colon tissues (n = 5 mice per group). i mRNA level of Ripk1 was analyzed by RT-PCR in HDL-16-L, HDL-16-H or PPTN administrated experimental colitis mice intestinal epithelial cell (n = 5 mice per group). j ChIP with anti-CREB of the regions containing the CREB binding sites on the Ripk1 gene promoter in IECs of HDL-16-L, HDL-16-H or PPTN administrated experimental colitis mice (n = 3 mice per group). k PI positive cells were analyzed by PI/Hoechst staining in TNF-α adding Smac mimetic and z-VAD (TSZ) treated HT-29 cells after pre-administrated with HDL-16 (n = 3 samples per group). l The PI staining and quantification of intestinal organoids from P2Y₁₄R^fl/fl mice treated as indicated (n = 3 samples per group). The data represent the mean ± SD for c, e, and h–l, the data represent the mean ± SEM for b. The p-values were determined by One-way ANOVA with Tukey multiple comparison test for c, e, i–l or two-way ANOVA with Sidak’s multiple comparisons test for b and h. For d, f, and g, each image was acquired independently three times, with similar results. Source data are provided as a Source Data file.