Fig. 2. PRMT5 inhibits cGAS-triggered type I IFN production.
(A and B) Quantitative polymerase chain reaction (PCR) analysis of Ifnb1 (A), Cxcl10, and Mx1 (B) mRNA in mouse BMDMs pretreated with EPZ015666 (5 μM) for 48 hours followed by the indicated transfection/infection. (C and D) Enzyme-linked immunosorbent assay (ELISA) analysis of IFN-β (C) and cGAMP (D) production in EPZ015666-pretreated BMDMs followed by the indicated transfection/infection. (E and F) Mouse BMDMs were transfected with Prmt5-siRNA for 48 hours, and Ifnb1, Cxcl10, and Mx1 mRNA (E) and IFN-β production (F) after HSV-1 infection were analyzed. (G) Luciferase reporter activity analysis of IFN-β (left) and ISRE (right) in HEK293T cells with the indicated transfection. (H) Immunoblot analysis of cGAS signaling in the EPZ015666-pretreated (left) or Prmt5-siRNA–transfected (right) mouse BMDMs followed by HSV-1 infection. (I and J) The shRNA-Ctr or shRNA-Prmt5–transfected MEF cells were further transfected with PRMT5(WT) or PRMT5(E444Q). Ifb1 mRNA (I) and cGAS signaling (J) in these transfected cells was detected by real-time PCR and western blot, respectively. *P < 0.05, **P < 0.01, and ***P < 0.001 (n = 3 mice per group, and the dots represent the value of each mouse). UT, untreated group; NS, not significant (I). Data are representative of three independent experiments with similar results [means ± SD in (A) to (G) and (I)]. DMSO, dimethyl sulfoxide.