Fig. 5. Macrophage glycolysis induces macrophage activation.

(A) qRT-PCR analysis of gene expression in Pfkfb3^WT or Pfkfb3^ΔMϕ BMDMs exposed to VCBM or Hx-REC-CM for 12 hours (n = 4). *P < 0.05 versus VCBM_Pfkfb3^WT; #P < 0.05 versus Hx-REC-CM_Pfkfb3^WT. (B) qRT-PCR analysis of gene expression in macrophages/microglia isolated from RA or OIR retinas in Pfkfb3^WT or Pfkfb3^ΔMϕ mice at P17 (n = 4). *P < 0.05 versus Pfkfb3^WT_RA; #P < 0.05 versus Pfkfb3^WT_OIR. (C to H) Mutiple IHC of Arg1 (C), Il1β (D), Fgf2 (G), F4/80, and isolectin on the sections of P17 OIR retinas (n = 6 to 8). Fluorescence intensity of Arg1 (D), Il1β (F), and Fgf2 (H) staining was calculated by ImageJ software. ***P < 0.001 versus Pfkfb3^WT. (I to J) Representative flow cytometric plots of retinal CD11b⁺F4/80⁺ cells showed CD86⁺CD206⁺ fractions in the retinas of RA and OIR mice at P17 (I) and comparisons of CD86⁺CD206⁺ fractions between genotypes (J) (n = 5 to 6). *P < 0.05; ***P < 0.001 versus Pfkfb3^WT_RA; ###P < 0.001 versus Pfkfb3^WT_OIR. Data are means ± SEM. P value determined by Student’s t test [for (D), (F), and (H)] and one-way ANOVA followed by Bonferroni test [for (A), (B), and (J)].