Figure 5.
PRMT1V2 (P1V2) interacts with PGC1α to modulate thermogenic activation. (A–C) qPCR analyses of thermogenic markers after infection with adenoviral vectors overexpressing GFP, PRMT1V1 (P1V1), or P1V2 along with adenoviral delivery of transcriptional regulators including (A and B) PGC1α or (C) PRDM16 in differentiated mouse primary EMSCs isolated from Prmt1fl/flAQcre mice (n = 6 per group). (D) ODD-luciferase activity in differentiated mouse primary EMSCs coinfected with adenoviral vectors overexpressing PGC1α and GFP or P1V2 after transduction of adenoviral-OLTAM (n = 6 per group). (E) CoIP of P1V2 with PGC1α. 293T cells were transfected with plasmids as indicated. Lysates were immunoprecipitated with an anti-HA antibody. Both input and immunoprecipitates were analyzed by immunoblotting as indicated. (F and G) Ucp1 promoter activity in 293T cells transiently transfected with Ucp1 promoter luciferase-reporter plasmid and vectors encoding transcriptional regulators PPARγ, (F) PGC1α or (G) PRDM16, and empty vector or P1V2 as indicated (n = 6 per group). (H and I) qPCR analyses of thermogenic gene expression after coinfection with adenoviral vectors overexpressing PGC1α and GFP, (H) P1V2 or (I) P1V1 in differentiated human primary ASCs (n = 6 per group). Data are presented as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001. n.s., not significant (P > 0.05). IB, immunoblot.