Evaluating PHGDH Inhibitors on Cell Proliferation and Apoptosis

CBR-5884 (Focus Biomolecules, Plymouth Meeting, PA, USA) and NCT-503 (Aobious, Gloucester, MA, USA) were used as PHGDH inhibitors (13 (link),14 (link)). The chemical structures of these inhibitors are in Supplementary Figure 1B and 1C. Cell proliferation was determined with XTT assays (Roche Applied Science, Tokyo, Japan) according to the manufacturer’s instructions. Cell apoptosis assays were carried out by flow cytometry (CytoFLEX analyzer; Beckman Coulter, Brea, CA, USA) using a FITC Annexin V Apoptosis Detection Kit (BD Biosciences, Bedford, MA, USA) according to the manufacturer’s recommendations. As a positive control, we used 5 μg/mL cycloheximide (Sigma, St. Louis, MO, USA) in apoptosis assay. We performed colony formation assay as previously described (15 (link)).