Figure 2. STAT3 is dynamic during wound repair and regulates SETDB2 in human and murine wound macrophages.

(A) Gene structure of the human SETDB2 gene showing cloned fragment aligned with multiple active transcriptional features including DHS peaks, H3K27Ac, H3K4me3, and sequence conservation. (B) Dot plot of different STAT members expression from scRNA-Seq data of human wounds (n = 10 patients). (C) Scatterplot of SETDB2 and STAT3 expression in macrophages obtained from scRNA-Seq in human wounds with respective Pearson correlation analysis (n = 10 patients). (D) ChIP-qPCR for Stat3 binding at the mouse Setdb2 promoter compared with IgG negative control. (E) Luciferase activity of the 3 kb human SETDB2 promoter cloned into pGL3 and then transfected in BMDMs untreated or treated with IFN-β (10 U/mL; 8.5 ng/mL), or IFN-β plus tofacitinib (100 μM) for 4 hours. (F) qPCR analysis of Setdb2 expression in wound macrophages (CD3^–CD19^–NK1.1^–Ly6G^–CD11b⁺) isolated from Stat3^fl/fl Lyz2^Cre mice on day 5 after wounding compared with Cre^– littermate control (n = 6 mice per group). (G) Setdb2 expression in BMDMs treated with IFN-β or IFN-β plus tofacitinib (n = 6–8 mice per group). (H) Representative Western blot and densitometry of murine whole wounds showing decreased levels of STAT3 at day 5 compared with day 0 after wounding (n = 3–4 mice at each time point). (I) Wound curve analysis in Stat3^fl/fl Lyz2^Cre mice compared with Cre^– littermate controls all fed a normal diet (n = 8–12 mice per group in each experiment). All data are representative of n = 3–5 independent experiments. *P < 0.05, **P < 0.01, ****P < 0.0001. Data are presented as the mean ± SEM. Two-tailed Student’s t test was used for comparison of 2 groups. For comparison among multiple groups, 2-way ANOVA followed by Newman-Keuls post hoc test was used.