Figure 2. Adoptive transfer of Sirt3-expressing macrophages decreases inflammation and improves wound healing in Sirt3-deficient mice.

A: Monocyte/macrophage (CD11b⁺CD3⁻CD11c⁻CD19⁻Ly6G⁻NK1.1⁻) single cell suspensions were isolated from Sirt3^−/− and Sirt3^+/+ spleens by cell sorting. 1 × 10⁶ cells were injected intravenously into wounded Sirt3^−/− mice. Wound closure was measured using ImageJ software (n=5 mice, repeated 1x). B: Monocyte/macrophage (CD11b⁺CD3⁻CD11c⁻CD19⁻Ly6G⁻NK1.1⁻) single cell suspensions were isolated from B6.SJL-PtprcaPepcb/BoyJ spleens by cell sorting. 1 × 10⁶ cells were injected intravenously into wounded Sirt3^−/− mice. Wounds were harvested and processed for CD11b⁺CD3⁻ CD19⁻Ly6G⁻ 3 days post-injury and analyzed for CD45.1 and CD45.2 surface marker expression by flow cytometry. Bar chart shows relative levels of CD45.1 and CD45.2 as a percent of CD11b⁺CD3⁻ CD19⁻Ly6G⁻ cells (n=3 mice). C: Monocyte/macrophage (CD11b⁺CD3⁻CD11c⁻CD19⁻Ly6G⁻NK1.1⁻) single cell suspensions were isolated from Sirt3^+/+ or Sirt3^−/− spleens by cell sorting. 1 × 10⁶ cells were injected intravenously into Sirt3^−/− mice and wounded. Wounds were harvested and processed for CD11b⁺CD3⁻ CD19⁻Ly6G⁻ 3 days post-injury. Relative gene expression of Il1b, Tnfa, and Nos2 was measured (n=6 mice). Data are presented as mean ± SEM. Data were analyzed for normality and 2-test Student t-test was performed. For data with multiple comparisons, ANOVA followed by Newman-Keuls multiple comparisons test was performed.