Fig. 4

GPR65 promotes M1 macrophage polarization in vitro. a qRT-PCR was used to assess the mRNA level of Gpr65, Ccl2, Tnfα, Il1β, Il6, Ccl5, Ccr2, Nos2, Cd80, Cd86, Mrc1, Arg1, Cd163 and Il10 in HMs isolated from WT and GPR65-KO mice (n = 4). b The expression and location of TNF-α in HMs isolated from WT and GPR65-KO mice were assessed by confocal microscopy. Scale bar = 20 μm. c TNF-α and IL-6 level in the supernatant of HMs isolated from WT and GPR65-KO mice were detected by ELISA (n = 4). HMs were transfected with pcDNA3.1 or pcDNA3.1-GPR65 for 48 h, qRT-PCR was used to assess the mRNA level of Gpr65, Ccl2, Tnfα, Il1β, Il6, Ccl5, Ccr2, Nos2, Cd80, Cd86, Mrc1, Arg1, Cd163 and Il10 (n = 4) (d); the expression and location of TNF-α was assessed by confocal microscopy (e). Scale bar = 20 μm. TNF-α and IL-6 level in the supernatant were detected by ELISA (n = 4) (f). ^*P < 0.05 vs. WT or pcDNA3.1. ELISA enzyme-linked immunosorbent assay, HM hepatic macrophage, KO knockout, qRT-PCR quantitative real-time reverse transcription-polymerase chain reaction, TNF-α tumor necrosis factor-α, IL-6 interleukin-6, n.d. not detected