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. Author manuscript; available in PMC: 2020 Dec 21.
Published in final edited form as: Sci Transl Med. 2020 Aug 5;12(555):eaay1371. doi: 10.1126/scitranslmed.aay1371

Fig. 4. EC-secreting soluble factor(s) induce macrophage glycolysis and polarize cells to mixed M1/M2-like macrophages.

Fig. 4.

(A) qRT-PCR analysis of the mRNA expression of glycolytic genes in BMDMs exposed to VCBM, Nx-REC-CM, or Hx-REC-CM for 12 hours (n = 4). *P < 0.05 versus VCBM. (B and C) Western blot analysis of Pfkfb3 protein expression in BMDMs exposed to VCBM, Nx-REC-CM, or Hx-REC-CM for 6 hours (n = 6). (D) The amount of intracellular lactate of BMDMs exposed to VCBM, Nx-REC-CM, or Hx-REC-CM for 12 hours. n = 5. **P < 0.01; ***P < 0.001. (E and F) Flow cytometry analysis of mixed M1/M2-like macrophages (CD11b+, F4/80+, CD86+, and CD206+) in the retinas of RA and OIR mice at different time points from P12 to P25 n = 4). *P < 0.001 and ***P < 0.001 versus RA. Data are means ± SD. (G) Heatmap displaying the fold changes of gene expression detected by qRT-PCR in macrophages/microglia isolated from RA or OIR retinas in WT mice at P17 (n = 4). (H) qRT-PCR analysis of gene expression in BMDMs exposed to VCBM, Nx-REC-CM, or Hx-REC-CM for 12 hours (n = 4). *P < 0.05 versus VCBM; #P < 0.05 versus Nx-REC-CM. Data are means ± SEM. P value determined by one-way [for (A), (C), (D), and (H)] or two-way [for (E)] ANOVA followed by Bonferroni test.

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