Fig. 6

LBP activates the TLR4/NF-κB pathway in intrahepatic macrophages to promote TGF-β1 secretion, in turn, TGF-β1 activates HSCs to direct intrahepatic fibrotic PMN. A Representative image of silver staining showed the proteins that were pulled down by immunoprecipitation of LBP overexpression in THP-1 cells. B Representative secondary mass spectrum of TLR4 protein. C Representative IB images of CO-IP with LBP or TLR4 overexpression in THP1cells. D Volcano plot of mRNA sequencing in PMA-treated THP-1 cells with IgG vs reLBP pretreatment in vitro. E ELISA was applied to determine TGF-β1 concentration in the supernatant of PMA-treated THP-1 cells with PBS, IgG or reLBP education, respectively. F IF staining was performed to investigate the regulatory mechanism of LBP in THP1 in vitro. G Representative IF images of TGF-β1 secretion in macrophages in the livers of mice with PBS, IgG or reLBP education, respectively. H-I IF staining (H) and WB (I) were performed to conform that the CM from reLBP-pretreated THP-1 cells activated LX-2 to increase the markers of fibrosis, whereas the anti-TGF-β1 antibody and galunisertib inhibited TGF-β/Smad signaling pathway to block the activation of LX-2 induced by the CM from reLBP-treated THP-1 cells. Data are representative of three independent experiments. Data pooled as mean ± SD of 5 independent experiments (C), and p values were determined by one-way ANOVA test (* P < 0.05, ** P < 0.01, *** P < 0.001)