Fig. 1

Jmjd3 directs early inflammation in nondiabetic wound Mφs via the JAK1/3/STAT3 pathway. A Jmjd3 expression from wound Mφs (CD3^-/CD19^-/NK1.1^-/Ly6G^-/CD11b⁺) harvested on Days 1–10 after wounding (6 mm punch biopsy; N = 4/group, repeated two times). B Tnfa and Il1b expression measured in wound Mφs isolated on Day 3 post-injury from Jmjd3^f/fLyz2^Cre+ mice and littermate controls (N = 5/group, pooled and repeated in triplicate). C Protein levels of TNF-α and IL-1β analyzed by ELISAs from Jmjd3^f/fLyz2^Cre+ wound Mφs and controls isolated on Day 3 post-injury (N = 5/group, pooled and repeated in triplicate). D Wound Mφs from Jmjd3^f/fLyz2^Cre+ mice and littermate controls were isolated on Day 3, and chromatin immunoprecipitation (ChIP) analysis for H3K7me3 on the Tnfa and Il1b promoters was performed compared to the IgG control (dotted line). (N = 4/group, pooled and repeated in triplicate). E Wound Mφs were isolated on Day 3 post-injury from Ifnar^−/− mice and controls (Ifnar^+/+) and stimulated ex vivo with IFN-β (100 U; 8.5 ng/mL) for 6 h, and Jmjd3 expression was analyzed by RT‒PCR (N = 3–5/group, pooled, repeated in triplicate). F ChIP analysis of H3K27me3 at the Il1b and Tnfa promoters from Day 3 isolated wound Mφs following ex vivo IFN-β stimulation (100 U; 8.5 ng/mL) for 6 h. G Jmjd3 expression in wound Mφs following ex vivo IFN-β stimulation (100 U; 8.5 ng/mL) with and without JAK1,3 inhibition with tofacitinib (100 μM; 6 hr incubation, N = 5/group, pooled, repeated in triplicate). H ChIP analysis for H3K27me3 at the Il1b and Tnfa promoters in wound Mφs following ex vivo IFN-β stimulation (100 U; 8.5 ng/mL) with and without JAK1,3 inhibition with tofacitinib (100 μM; 6 hr incubation, N = 5/group, pooled, repeated in triplicate). I Jmjd3 expression in Stat3^f/fLyz2^Cre+ mice and littermate control wound Mφs following ex vivo IFN-β stimulation (100 U; 8.5 ng/mL, N = 5/group, repeated in triplicate). H ChIP analysis of H3K27me3 at the Il1b and Tnfa promoters in Stat3^f/fLyz2^Cre+ wound Mφs following ex vivo IFN-β stimulation (100 U; 8.5 ng/mL) for 6 h (N = 5/group, pooled, repeated in triplicate). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Data are presented as the mean ± SEM. Data were first analyzed for a normal distribution, and if the data passed the normality test, a two-tailed Student’s t test was used