Extended Data Fig. 7. Utility of Enolase inhibitors beyond cancers with ENO1-deletions.

Hyperactivation of HIF by loss of VHL tumor suppressor gene has been reported to sensitize cells to glycolysis inhibition¹². a. Proliferation of renal clear cell carcinoma cell lines that lack functional VHL (786-O and RCC4, red) and isogenic rescued cell lines re-expressing VHL (blue) was followed by Incucyte live cell imager (x-axis, time; y-cell confluence) in response to POMHEX treatment. Each box represents one biological replicates. VHL-deleted parental lines showed 4- to 8-fold greater sensitivity than isogenic rescued VHL lines. b. Cell density quantified by crystal violet for the same cell lines corroborates Incucyte live imaging data (N = 4 biological replicates, with +/− S.E.M.). IC₅₀ for POMHEX in the RCC4 line is ~250 nM and 1,000 nM in the isogenic rescued line. TCA-cycle deficiency and defective oxidative phosphorylation by loss of function of Fumarase (FH) increase reliance on glycolysis and sensitize to glucose deprivation¹¹. c. Likewise, the FH-null renal carcinoma cell line, UOK262, (red) shows dramatically higher sensitivity to the Enolase inhibitor POMHEX than isogenic FH-rescued cell line control (blue). Cell density in response to POMHEX treatment was quantified by crystal violet (N = 4 biological replicates, with +/− S.E.M.).