Fig. 5

IL-33 improves the healing outcome and modulated the immune microenvironment in RISI. A Schematic representation of subcutaneous injection of IL-33 neutralizing antibody or control IgG. B Representative images of gross injury after IL-33 neutralizing antibody injection compared to control IgG injection, n = 3. C Gross injury scores for the IL-33 neutralizing antibody injection group compared to the control groups, n = 3 for each group. D Representative H&E staining images of IL-33 neutralizing antibody injection groups versus control groups. Representative immunofluorescence images of E Ki67⁺ proliferating cells, F α-SMA⁺ myofibroblasts, G CD31⁺ endothelial cells and H Cd11b⁺ myeloid immune cells for the IL-33 neutralizing antibody injection groups versus the control groups. I Quantitative analysis of Ki67, α-SMA, and CD31 positive signals in the IL-33 neutralizing antibody injection groups versus the control groups. Twelve high-power fields were randomly selected, and positive signal cell ratios (for Ki67) or positive signal areas (for α-SMA and CD31) were analyzed using ImageJ software. J qPCR analysis of changes in M2 macrophage marker CD206 and M1 macrophage marker CD86 gene expressions after IL-33 neutralizing antibody injection compared to control groups, n = 6. K Western blot indicated a decrease in the anti-inflammatory cytokine Tgf-β expression level and an increase in the pro-inflammatory cytokine IL-6 expression level after IL-33 neutralizing antibody injection, with three repetitions, and representative images was shown. Data were represented as mean ± standard deviation. When variance was met, a t-test was used for statistical analysis between two groups, while ANOVA was used for comparisons among three or more groups. If variance wasn’t met, the Mann-Whitney U test was used. *p < 0.05, **p < 0.01, ***p < 0.001. Bars represented 100 μm