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. Author manuscript; available in PMC: 2021 Mar 1.
Published in final edited form as: J Bone Miner Res. 2019 Dec 12;35(3):550–570. doi: 10.1002/jbmr.3908

Figure 7: Inhibition of H+/lactate Efflux Decreases ECAR without Increasing Mitochondrial OCR; Resulting intracellular lactate accumulation increases HIF-1α activity.

Figure 7:

A- B’) Seahorse Flux analysis of extracellular acidification rate (ECAR) over time (A, B) and on average (A’, B’) after MCT inhibition with an MCT inhibitor cocktail (MCT Inh) (A, A’) and Syrosingopine (Syro) (B, B’). C) MCT inhibition with MCT Inh. and Syro decreases extracellular lactate concentrations. D) Intracellular pH measurement after MCT inhibition with MCT Inh. and Syro. E) NAD+/NADH ratio after MCT inhibition. F-G’) Seahorse Flux analysis of oxygen consumption rate (OCR) over time (F, G) and on average (F’, G’) after MCT inhibition with MCT Inh (F, F’) and Syro (G, G’). H) Schematic of HIF-1α degradation vs. stabilization depending on status of PHD function. I, J) 5 mM exogenous lactate (I) and MCT inhibition (J) increase HIF activity as measured by dual luciferase assay. n= 4 independent experiments; 4-6 replicates/experiment. Statistical analysis: t-test, One-way ANOVA. n.s.= not significant; *, p-value ≤ 0.05; **, p-value ≤ 0.01; ***, p-value ≤ 0.001.