Figure 3. STAT3 inhibits the physical interaction between SETDB2 and NF-κB in wound macrophages.

(A) Heatmap of Pearson gene correlation analysis results for SETDB2, STAT3, C-REL, and RELA in macrophages obtained from scRNA-Seq in human wounds (n = 10 patients). (B) Western blot showing Setdb2-interacting proteins purified after incubating GST-SETDB2 with BMDM lysate. (C) Western blot of RelA coimmunoprecipitated with Setdb2 from BMDMs treated with Stat3 inhibitor compared with untreated BMDMs. (D) Setdb2 immunoprecipitate reactions from BMDMs isolated from Stat3^fl/fl Lyz2^Cre+ mice and Cre-negative littermate controls that were untreated or treated with IFN-β (10 U/mL; 8.5 ng/mL) for 4 hours were analyzed via Western blot and probed for RelA and Setdb2 (n = 4–6 mice per group). (E and F) ChIP-qPCR on BMDMs isolated from Stat3^fl/fl Lyz2^Cre+ versus Cre-negative controls for NF-κB (RelA) and Setdb2 at the Il1b and Il6 promoters (n = 4–6 mice per group). All data are representative of n = 3–5 independent experiments. **P < 0.01. 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.