Fig 1: Effect of PRMT5 inhibitor on macrophage activation. THP-1 cells were pretreated with phorbol 12-myristate 13-acetate and then treated with PRMT5 inhibitor EPZ015666 (10 µM) for 24 h, followed by incubation with RPMI-1640 containing homogenized solution of normal and eutopic endometrium (100 µl/ml) for additional 3 days. ELISAs were performed to determine the concentrations of IL-6 and IP-10 in the medium. **P<0.01 (n=15). PRMT5, protein arginine methyltransferase 5; IP-10, IFN-?-induced protein 10.
Fig 2: Schematic diagram illustrating how the eutopic endometrial microenvironment regulates macrophage PRMT5 expression via NF-?B signaling. PRMT5, protein arginine methyltransferase 5; IP-10, IFN-?-induced protein 10.
Fig 3: A proposed working model of the role of the ZEB2/TWIST1/PRMT5/NuRD complex in EMT and tumor metastasis in CRC.
Fig 4: Effect of extracts of eutopic endometrium from patients with endometriosis on macrophage PRMT5 expression. THP-1 cells were pretreated with phorbol 12-myristate 13-acetate and then stimulated with 100 ng/ml LPS for 24 h, followed by incubation with RPMI-1640 containing homogenized solution of normal and eutopic endometrium (100 µl/ml) for 3 days. Reverse transcription-quantitative PCR and western blot analyses were performed to examine the mRNA and protein expression levels of PRMT5 in THP-1 cells. ß-actin was used as internal control. *P<0.05; **P<0.01 (n=25). PRMT5, protein arginine methyltransferase 5; LPS, lipopolysaccharide.
Fig 5: 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-ß (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-ß production (F) after HSV-1 infection were analyzed. (G) Luciferase reporter activity analysis of IFN-ß (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.
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