Fig 1: Fluorometric NADH-linked assay for measuring Enolase activity.The effect on ENOblock (a, b) and SF2312 (b, d) on Enolase activity in lysates from the D423 cell line expressing ENO2 (red diamonds) or ENO1 (blue squares) was determined using the NADH-linked assay. Panel b shows a representative trace of NADH fluorescence over time of vehicle control (gray symbols) and 100 μM ENOblock (orange symbols), while Panel d shows traces of vehicle control (gray symbols), with 50 nM SF2312 (red symbols). Panel a and c shows enolase activity normalized to vehicle control and expressed as a function of inhibitor concentration. Each data point represents mean of N = 6 (a) and N = 4 (c) ± S.D.
Fig 2: Non-selective toxicity of ENOBlock to ENO1-deleted glioma cells.A representative plate of cancer cells treated with ENOblock is shown in panel a, with quantification shown in panel b A plate treated with SF2312 is shown in panel c, with quantification shown in panel d. Cell were treated for 7 days. (b, d) D423 ENO1-deleted (red diamonds), D423 ENO1-rescued (blue squares) and LN319 ENO1 WT (grey circles) were treated with the indicated doses of ENOblock in panel b (N = 4 ± S.D) or SF2312 in panel d (N = 4 ± S.D). Cell density was quantified by crystal violet and expressed relative to vehicle control as a function of inhibitor concentration. At high concentrations, SF2312 selectively killed D423 ENO1-deleted cells as compared to D423 ENO1-rescued cells (p<0.05, Repeated Measures one-way ANOVA with Bonferroni correction). ENOblock failed to show such selectivity regardless of dose.
Supplier Page from Abcam for Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407]