Validation of NLRP3 and NLRP1 in the RPE cells of EAU with IFN-λ deficiency (IMAGE)
Caption
(A, B) The primary RPE cells from IFNLR1−/− EAU mice and WT EAU mice were harvested for the detection of NLRP3 and NLRP1. (A) The mRNA expression of NLRP3 and NLRP1 was detected by RT-qPCR. (B) The representative Western blotting images and histogram (n = 4). (C, D) Primary hRPE cells were transfected with siIFNLR1, and then the cells were assayed for NLRP1 and NLRP3 expression by RT-qPCR and Western blotting (n = 4). (E) Primary RPE cells from IFNLR1−/− EAU mice and hRPE cells with siIFNLR1 were treated with or without MCC950 (a potent, selective and small-molecule inhibitor of NLRP3) or DPP9 (a direct inhibitor of NLRP1) in the presence of LPS stimulation, and the supernatants were harvested for the expression of inflammatory cytokines including IL-6, IL-8, MCP-1, IL-1β and TNF-α by ELISA (n = 5). ∗∗P < 0.01, ∗∗∗P < 0.001; P value was corrected by Bonferroni. EAU, experimental autoimmune uveitis; ELISA, enzyme-linked immunosorbent assay; LPS, lipopolysaccharide; RPE, retinal pigment epithelium; RT-qPCR, real-time quantitative PCR.
Credit
Wujiao Wang, Qingfeng Wang, Jinyu Cai, Guannan Su, Wanyun Zhang, Shuai Su, Yunfan Zheng, Chaokui Wang, Peizeng Yang
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