Fig. 4: Glyoxalase I is required for methylglyoxal detoxification.
From: Reactive metabolite production is a targetable liability of glycolytic metabolism in lung cancer
a Western blot analysis of glyoxalase I (Glo1) expression in parental 3553T3 lung cancer cells and in three independent clones where the Glo1 gene was disrupted using CRISPR/Cas9 (sgGlo1). b Relative levels of lactoylglutathione (LGSH) in 3553T3 parental NSCLC cells or the Glo1-deleted clones described in (a) as detected using LCMS. Values shown are the peak area of LGSH normalized to the peak area of GSH (n = 3). c Relative number of viable 3553T3 parental cells and Glo1-deleted clones as determined by CellTiter-Glo Luminescence Assay following a 48-h incubation with 200 μM methylglyoxal (n = 3). d LCMS quantification of relative free MG-H1 abundance in 3553T3 parental cells and Glo1-deleted clones. Values are peak area of MG-H1 normalized to peak area of arginine (n = 3). e LCMS quantification of free MG-H1 abundance in extracts from 3553T3 parental cells and Glo1-deleted clones that were incubation with 200 μM methylglyoxal for 16 h. Values are normalized to MG-H1 peak area in parental lung cancer cell lines incubated without methylglyoxal treatment shown in panel (d), which were analyzed in the same LCMS experiment (n = 3). f Western blot analysis using an antibody raised against the MG-H1 epitope of lysates from 3553T3 parental cells and Glo1-deleted clones that had been treated without or with 200 μM of methylglyoxal for 12 h. Values in b–e denote mean ± SEM and the P values were calculated by unpaired, two-tailed t-test.
