Supplementary Materials

The PDF file includes:

  • Materials and Methods
  • Fig. S1. Retinal macrophages/microglia are hyperglycolytic in the pathological angiogenic vascular niche in a mouse model of OIR.
  • Fig. S2. Generation of myeloid-specific Pfkfb3 KO mice.
  • Fig. S3. Effect of myeloid Pfkfb3 deficiency on the development of retinal vasculature in mice.
  • Fig. S4. Effect of myeloid Pfkfb3 or Pkm2 deficiency on retinal vaso-obliteration at P12 in OIR.
  • Fig. S5. Effect of hypoxic REC-CM on glycolytic function and mitochondrial respiration function of BMDMs.
  • Fig. S6. Effect of hypoxic REC-CM on retinal microglia glycolysis and polarization.
  • Fig. S7. Macrophages/microglia exhibit mixed M1/M2-like phenotype in OIR retinas.
  • Fig. S8. Expression of M1/M2 markers in retinal macrophages/microglia in OIR retinas at different time points.
  • Fig. S9. Immunofluorescent staining shows the colocalization and expression of CD11c and CD206 within the pathological neovascular tufts of OIR retina at P17.
  • Fig. S10. Mixed M1/M2 population of macrophages is not found in adipose tissue from mice fed an HFD.
  • Fig. S11. Hypoxia alone is not sufficient to cause a mixed M1/M2-like phenotype in macrophages.
  • Fig. S12. Effect of glycolysis on macrophage M1/M2 polarization.
  • Fig. S13. Effect of Pfkfb3 KO on IL-4–induced macrophage M2 polarization.
  • Fig. S14. Effect of Pfkfb3 KO on the number of macrophages and neutrophils in RA or OIR retinas.
  • Fig. S15. Effect of Pfkfb3 KO on macrophage proliferation and viability.
  • Fig. S16. Hx-REC-CM–treated macrophages promote EC sprouting though increased secretion of proinflammatory and proangiogenic factors.
  • Fig. S17. REC-secreting metabolites are sufficient to induce macrophage glycolysis and activation.
  • Fig. S18. Effect of acidic pH on macrophage activation.
  • Fig. S19. Effect of histone acetylation inhibition on the Hx-REC-CM–induced expression of macrophage polarization markers and proangiogenic factors.
  • Fig. S20. Schematic diagram illustrating the proposed mechanisms for PFKFB3-driven glycolysis-mediated reciprocal activation between macrophages/microglia and ECs in the retinal angiogenic niche.
  • Table S1. Primer sets used for qRT-PCR.
  • Table S2. Primer sets used for ChIP.
  • References (4649)

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