FIGURE 4.

Effects of Ag120 and glutamine depletion on CRC cells. (A,B) 2-HG production (A) and immunoblot analysis of IDH1mt expression (B) in CRC cells treated with Ag120 or V9302 for 72 h. (C) Quantification of viable HCT116 and HT29 cells treated with vehicle or Ag120 (45 µM) for 72 h in DMEM medium containing 2 or 0 mM L-glutamine. Cell numbers were measured using a CCK-8 kit. Contribution of Ag120 to cell viability was calculated based on the data in Supplementary Figure S3A. (D) Colony formation by CRC cells after 10 days of treatment with 0 or 45 µM Ag120 in 0 or 2 mM L-glutamine-containing medium. (E,F) Fluorescence microscopy images of EdU incorporation in HCT116 (E) and HT29 (F) cells treated for 72 h with 0 or 45 µM ASCT2 in 0 or 2 mM L-glutamine-containing medium. Scale bars, 50 µm. Mean ± SD of three experiments. *p < 0.05, **p < 0.01 by Student’s t-test.