EDG5 Antibody - #DF4921

Figure 5 Systemic Mas1 deficiency inhibits S1P-induced NETs formation. A) GSEA of GO enrichment plot in livers from WT-L/G and Mas1−/−-L/G mice (both n = 3, bulk RNA-seq). B) PCA of liver metabolomics (n = 6 per group, left) and the relative abundance of intrahepatic S1P (right) (two-sided Mann–Whitney U test, p = 2.58 × 10−2). C) Serum S1P levels (all, n = 6, One-way ANOVA with Tukey's test, two-sided Student's t-test, p = 4.28 × 10−4 and p = 3.27 × 10−4 from left to right). D) Intrahepatic mRNA expression of Sphk1 and S1pr2 (all, n = 6, One-way ANOVA with Tukey's test, two-sided Student's t-test, p = 3.12 × 10−4, p = 5.06 × 10−4, p = 4.18 × 10−4 and p = 3.91 × 10−4 from left to right). E) Representative immunoblots of liver tissues with the quantification (below). F) The violin boxplot (left) illustrates S1pr2 expression across cell types, and the dot plot (right) presents S1pr1 through S1pr5 expression in neutrophils (scRNA-seq). G) The mRNA expression of S1pr2 in hepatic neutrophils (both n = 6, two-sided Student's t-test, p = 8.12 × 10−3). WT-L/G mice were pre-treated with Ang-(1-7) or solvent control (n = 6 per group, H–J). H) Serum S1P levels (two-sided Mann–Whitney U test, p = 4.08 × 10−4). I) Intrahepatic mRNA expression of Sphk1 and S1pr2 (two-sided Student's t-test, p = 8.17 × 10−4 and p = 6.35 × 10−4 from left to right). J) Representative immunoblots of liver tissues with the quantification (below). WT-L/G mice were pre-treated with PF543 (SphK1 inhibitor) or solvent control (n = 6 per group, K–P). K) Representative liver photographs and immunohistochemical staining with the quantification (below) of H&E, TUNEL, and Ly6G (two-sided Student's t-test, p = 5.60 × 10−4, p = 3.19 × 10−4 and p = 8.72 × 10−4 from left to right). Scale bars are shown as indicated. L) Serum levels of ALT and TBA (two-sided Student's t-test, p = 3.96 × 10−4 and p = 4.18 × 10−4 from left to right). M) Serum levels of S1P (two-sided Student's t-test, p = 2.98 × 10−4). N) Intrahepatic mRNA expression of Sphk1 and S1pr2 (two-sided Student's t-test, p = 7.61 × 10−4 and p = 3.59 × 10−4 from left to right). O) Plasma levels of cell-free DNA (two-sided Mann–Whitney U test, p = 3.18 × 10−3). P) Representative immunoblots of liver tissues with the quantification (below). Data are presented as mean ± SD or median ± IQR (*p < 0.05, **p < 0.01, ***p < 0.001). See also related Figure S7 (Supporting Information).

Figure 1: Serum bile acid profile and intestinal flora were changed after CBDL. (A) Successful establishment of models for cholestasis-induced liver fibrosis. Representative images of H&E staining between sham group and CBDL group after 3 weeks of operation (scale bar = 50 um). (B) Serum targeted bile acids indicate a significant increase in conjugated bile acids in the CBDL group, compared to the sham group. (C) Immunohistochemistry results demonstrate a significant increase in S1PR2 expression in the CBDL group (scare bar = 20 um). (D) Semi-quantitative analysis for S1PR2 OD value. (E) Western blot results indicate a significant increase in liver S1PR2 expression in the CBDL group. (F) Quantitative analysis for S1PR2 protein. (G) The α-diversity of gut microbiota was determined using Chao, Sob, and Simpson indices (sham group; n = 12; CBDL group, n = 13). (H) Unweighted UniFrac-based principal coordinate analysis (PCoA) based on the ASV levels (sham group; n = 12; CBDL group, n = 13). (I) Microbial composition analysis at the phylum level in two groups of samples. (J) Differential microbiota between the sham group and the CBDL 3w group. CA = Cholic acid; CDCA = Chenodeoxycholic acid; TCA = Taurocholic acid; TCDCA = Taurochenodeoxycholic acid; DCA = Deoxycholic acid; LCA = Lithocholic acid; HCA = hyocholic acid. *P < 0.05, **P < 0.01, ***P < 0.001.

Figure 1: Serum bile acid profile and intestinal flora were changed after CBDL. (A) Successful establishment of models for cholestasis-induced liver fibrosis. Representative images of H&E staining between sham group and CBDL group after 3 weeks of operation (scale bar = 50 um). (B) Serum targeted bile acids indicate a significant increase in conjugated bile acids in the CBDL group, compared to the sham group. (C) Immunohistochemistry results demonstrate a significant increase in S1PR2 expression in the CBDL group (scare bar = 20 um). (D) Semi-quantitative analysis for S1PR2 OD value. (E) Western blot results indicate a significant increase in liver S1PR2 expression in the CBDL group. (F) Quantitative analysis for S1PR2 protein. (G) The α-diversity of gut microbiota was determined using Chao, Sob, and Simpson indices (sham group; n = 12; CBDL group, n = 13). (H) Unweighted UniFrac-based principal coordinate analysis (PCoA) based on the ASV levels (sham group; n = 12; CBDL group, n = 13). (I) Microbial composition analysis at the phylum level in two groups of samples. (J) Differential microbiota between the sham group and the CBDL 3w group. CA = Cholic acid; CDCA = Chenodeoxycholic acid; TCA = Taurocholic acid; TCDCA = Taurochenodeoxycholic acid; DCA = Deoxycholic acid; LCA = Lithocholic acid; HCA = hyocholic acid. *P < 0.05, **P < 0.01, ***P < 0.001.