Fig. 2
Both PRMT5 and PRMT1 interact with CFLARL. a 293FT cells were transfected with the pEBG and pEBG-CFLARL plasmids. The cells were harvested and prepared for the GST pull-down assay after 24 h, and the CFLARL protein was detected by western blotting. b Cells were transfected with the pcDNA3.1 and pcDNA3.1-MYC-PRMT5 plasmids. The cells were harvested and prepared for the immunoprecipitation assay after 24 h. The lysate supernatant was incubated with the MYC antibody for 1 h, followed by direct incubation with protein-A (1:1 mix) beads at 4 °C overnight. The precipitated proteins were analyzed by a western blot assay. c The same as described in (A). d Cells were transfected with the pcDNA3.1 and pcDNA3.1-MYC-PRMT1 plasmids. The cells were harvested and prepared for the immunoprecipitation assay after 24 h. The lysate supernatant was incubated with the MYC antibody for 1 h and then directly with protein-A beads at 4 °C overnight. The precipitated proteins were analyzed by western blot analysis. GAPDH was used as a loading control. e IP assay using CFLARL antibody was conducted in A549 cells and endogenous PRMT1/5 was detected