Fig. 2. PRMT1 catalyzes PGK1-ADMA at the R206 residue.

A–E HEK293T and HCT116 cells were transfected with exogenous Flag-PGK1 (A, B). Anti-FLAG beads were used for immunoprecipitation to detect the mutual binding between PRMT1 and PGK1, and the methylation changes of ADMA were also detected. Flag-PGK1 overexpressed HCT116 cells were transfected with exogenous Myc-PRMT1 (C), shPRMT1 (D), or treated with GSK3368715 (E) to detect the methylation changes of ADMA. F, G Methylation of arginine at R206 and R330 was detected by LC-MS. H, I Wild and mutant Flag-PGK1 (WT, R206K, and R330K) plasmids were overexpressed, and methylation changes of ADMA were detected by IP of Flag in HCT116 and HEK293T cells. The relative intensity of ADMA level were quantified by software Image J. The working concentration of GSK3368715 is 3.1 nM for 24 h.