Figure 4. Loss of AKT3 in M2 macrophages inhibited extracellular COL1A1 and COL11A1 expression. (A) GSEA showed that negatively enriched genes were associated with PI3K-AKT signaling and phagosomes in delayed cutaneous wound tissue. (B) Heatmap of the top 10 genes related to PI3K-AKT signaling and phagosomes; AKT3 was downregulated in both functional enrichment sets in the delayed cutaneous wound tissue. (C) Immunofluorescence of cutaneous wound tissue (n = 6). CD68- (green) and CD206-(red) positive M2 macrophages were reduced in the delayed cutaneous wound tissue. AKT3 (pink) was decreased in the M2 macrophages. (D) qRT-PCR showed decreased AKT3 mRNA expression in the delayed cutaneous wound tissue-derived M2 macrophages. (E) Western blotting verified the reduction and loss of AKT3 in M2 macrophages from delayed cutaneous wound tissue. (F) Immunofluorescence of COL1A1 and COL11A1 in CD68-positive macrophages in cutaneous wound tissue. (a) Decreased CD68-positive macrophage infiltration and COL1A1 protein expression were observed in delayed cutaneous wound tissue. (b) Decreased COL11A1 protein expression also accompanied the reduced CD68-positive macrophage infiltration. All the experiments were repeated at least three times.