Figure 2
Roxadustat increases glycolysis in primary Müller glia. (a) Glycolysis, glycolytic capacity, and glycolytic reserve were calculated from the extracellular acidification rate (ECAR) in primary Müller glia exposed for 48 h to Roxadustat, as shown in (b). Müller glia exposed to Roxadustat engaged in significantly higher glycolysis as compared to control Müller glia (*p = 0.015); glycolytic capacity did not vary across the control and Roxa-treated cells. The Roxa-treated cells did exhibit significantly lower glycolytic reserve (*p = 0.02) than the control cells. (b) The moment-by-moment ECAR measures taken during the glycolytic stress test as glucose (Glc), then oligomycin, and finally, 2-deoxyglucose (2-DG) were sequentially added to the wells containing either control or Roxa-treated Müller glia. Glycolysis in the Roxa-treated cells was significantly higher than control cells (*p = 0.015). Graph shows example trace from a biological replicate. (c) Primary Müller glia subjected to a fuel flex test using glucose showed that the Roxa-treated Müller glia were no different from control in terms of their dependency on glucose or their flexibility towards glucose utilization, but they had significantly lower capacity than the control cells (*p = 0.028). Seahorse Bioanalyzer experiments were run with three separate primary Muller glial cell isolations, with 4–9 wells assessed per run for the data shown in a and c.
