The small molecules methotrexate (Selleckchem, S1210), hypoxanthine (Sigma, H9636), thymidine (Sigma, T1895), and sodium formate (Fisher Scientific, S648-500) were used. The dual SHMT1/2 inhibitor SHIN1 (Tocris, 6998) was used at the concentration of 10 µM. In total, 1 mM of sodium formate was used to demonstrate on-target effects of SHIN1. Glutamylcysteine synthetase inhibitor l-buthionine-sulfoximine (BSO; Sigma, 83730-53-4) was used at the concentration of 10 µM. Mitochondrial complex I inhibitor Piericidin A (Cayman, 15379) was used at the concentration of 0.1 µM. Brequinar (Cayman, 24445) was used at the concentration of 1 µm and 10 µM. Glucose-free media containing galactose was prepared by supplementing 25 mM galactose (Sigma, G5388) into glucose-free DMEM (#11966025, ThermoFisher) with 10% dialyzed FBS (#26400044, ThermoFisher). Cells were always treated with small molecules or special media 2 h prior to infection. Samples were harvested at 48 hpi.
Zhang Y., Guo R., Kim S.H., Shah H., Zhang S., Liang J.H., Fang Y., Gentili M., Leary C.N., Elledge S.J., Hung D.T., Mootha V.K, & Gewurz B.E. (2021). SARS-CoV-2 hijacks folate and one-carbon metabolism for viral replication. Nature Communications, 12, 1676.
+ Open protocol