Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Siglec-H -/- Plasmacytoid Dendritic Cells Protect Against Acute Liver Injury by Suppressing IFN-γ/Th1 Response and Promoting IL-21 + CD4 T Cells

doi: 10.1016/j.jcmgh.2024.101367

Figure Lengend Snippet: Loss of SH prevents ConA-induced IFN-γ/Stat1 and JNK signalings activation and enhances Stat3, IFN-I, and IL21 levels in the liver. mRNAs and proteins were isolated from WT and SH KO mice livers 12 hours after ConA injection for analysis. Intrahepatic expression levels of mRNA of ( A ) IL6, ( B ) TNF-α, ( C ) IFN-γ, ( D ) interferon regulatory factor 1 (IRF-1), ( E ) CXCL-10, ( F ) SOCS3, ( G ) IFN-β, and ( H ) IL21 analysis by real-time quantitative PCR. ( I-L ) Immunoblot detection and quantification of anti-phospho- Stat1, Stat3, JNK, and their corresponding total isoforms, and β-actin. Target proteins quantification was performed with ImageJ and presented as ratio phosphorylated over total isoforms. Bars in the scatter plots represent the median value. Statistical analysis was performed with 2-way ANOVA and Tukey post hoc test; ∗ P < .05; ∗∗ P < .005; ∗∗∗ P < .0005; ∗∗∗∗ P < .00005.

Article Snippet: Animals were administrated with ConA (15 mg/kg, intravenously) from Sigma Millipore, SAP-conjugated anti-BDCA2 antibody or pDC-dAb (10 μg/mouse, intraperitoneally) for pDCs depletion, InVivoMAb antimouse IL21R antibody (1 mg/kg, intraperitoneally) from BioXCell, and Stat3 inhibitor HJC-0416 (10 mg/kg, intraperitoneally) from MedChemExpress.

Techniques: Activation Assay, Isolation, Injection, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Siglec-H -/- Plasmacytoid Dendritic Cells Protect Against Acute Liver Injury by Suppressing IFN-γ/Th1 Response and Promoting IL-21 + CD4 T Cells

doi: 10.1016/j.jcmgh.2024.101367

Figure Lengend Snippet: Signaling pathways in ConA-induced acute liver injury in WT and SH KO mice. Proteins were isolated from WT and SH KO mice livers 12 hours after ConA injection for analysis. Immunoblot detection and quantification of anti-phospho- Stat1, Stat3, JNK, and their corresponding total isoforms, and β-actin. Target proteins quantification was performed with ImageJ and presented as ratio phosphorylated over total isoforms. Quantification data were combined and presented in Figure 5 .

Article Snippet: Animals were administrated with ConA (15 mg/kg, intravenously) from Sigma Millipore, SAP-conjugated anti-BDCA2 antibody or pDC-dAb (10 μg/mouse, intraperitoneally) for pDCs depletion, InVivoMAb antimouse IL21R antibody (1 mg/kg, intraperitoneally) from BioXCell, and Stat3 inhibitor HJC-0416 (10 mg/kg, intraperitoneally) from MedChemExpress.

Techniques: Protein-Protein interactions, Isolation, Injection, Western Blot

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Siglec-H -/- Plasmacytoid Dendritic Cells Protect Against Acute Liver Injury by Suppressing IFN-γ/Th1 Response and Promoting IL-21 + CD4 T Cells

doi: 10.1016/j.jcmgh.2024.101367

Figure Lengend Snippet: IL21R and Stat3 blockade restores ConA-induced acute liver injury in SH-deficient mice. ( A ) Experiment design: SH KO mice treated with either anti-IL21R antibody or HJC-0416 (Stat3 inhibitor) and terminated 6 hours after ConA administration. Enzyme-linked immunosorbent assay detection of ( B ) ALT and ( C ) HA in the blood at 6 hours post-ConA injection. ( D ) H&E staining ( top ), and detection of apoptotic cells ( green ) with nuclei ( blue ) by tunnel assay ( bottom ) in liver sections of anti-IL21R- and HJC-0416-treated SH KO at 6 hours post-ConA injection, and ( E ) ImageJ quantification of tunnel positive area. All images were acquired with ×20 magnification lens. ( F - H ) Immunoblot detection and quantification of anti-phospho Stat3 ( G ) and JNK ( H ), and their corresponding total isoforms. Target proteins quantification was performed with ImageJ and presented as ratio phosphorylated over total isoforms. Bars in the scatter plots and histograms represent the median value and the SEM respectively. Statistical analysis was performed with 2-tailed unpaired t test; ∗ P < .05; ∗∗ P < .005; ∗∗∗ P < .0005; ∗∗∗∗ P < .00005.

Article Snippet: Animals were administrated with ConA (15 mg/kg, intravenously) from Sigma Millipore, SAP-conjugated anti-BDCA2 antibody or pDC-dAb (10 μg/mouse, intraperitoneally) for pDCs depletion, InVivoMAb antimouse IL21R antibody (1 mg/kg, intraperitoneally) from BioXCell, and Stat3 inhibitor HJC-0416 (10 mg/kg, intraperitoneally) from MedChemExpress.

Techniques: Enzyme-linked Immunosorbent Assay, Injection, Staining, Western Blot