rat anti mouse mab f4 80  (Bio-Rad)

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: XBP1 Facilitating NF-κB-p65 Nuclear Translocation Promotes Macrophage-Originated Sterile Inflammation Via Regulating MT2 Transcription in the Ischemia/Reperfusion Liver

doi: 10.1016/j.jcmgh.2024.101402

Figure Lengend Snippet: Myeloid-specific XBP1 deficiency decreases inflammatory cell infiltration and proinflammatory mediators in ischemic livers. The IRI model mice were subjected to 90-minute ischemia, followed by 6-hour reperfusion. ( A, B ) Macrophage infiltration was analyzed by immunohistologic staining with antibodies against F4/80 or CD11b; F4/80 + or CD11b + cells were quantitated by counting numbers of positive cells/area, with the scale of 200 μm. ( C ) Neutrophil infiltration was analyzed by immunohistologic staining with antibodies against Ly6G; Ly6G + cells were quantitated by counting numbers of positive cells/area, with the scale of 200 μm. ( D ) Detection of TNF-α, IL1β, IL6, iNOS, and IL10 in ischemic livers using qRT-PCR. ( E ) Detection of TNF-α, IL6, and IL10 in serum by enzyme-linked immunosorbent assay. n = 6, ∗ P < .05 by Student t test.

Article Snippet: The Suzuki score, which consists of 3 aspects (hepatocyte necrosis, sinusoidal congestion, and ballooning degeneration), was rated on a scale of 1 to 4, as defined by Suzuki et al. Hepatic macrophages and neutrophils were analyzed by IHC staining and incubation with primary rat anti-mouse F4/80, CD11b and Ly6G mAbs (Cell Signaling Technology).

Techniques: Staining, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: XBP1 Facilitating NF-κB-p65 Nuclear Translocation Promotes Macrophage-Originated Sterile Inflammation Via Regulating MT2 Transcription in the Ischemia/Reperfusion Liver

doi: 10.1016/j.jcmgh.2024.101402

Figure Lengend Snippet: MT2 inhibition abolishes XBP1 M-KO -related anti-inflammatory features in vivo and in vitro. The IRI model mice were subjected to 90-minute ischemia, followed by 6-hour reperfusion. ( A ) XBP1 M-KO mice were injected with siRNA-MT2 by caudal vein before establishing hepatic IRI model. ( B ) Western blotting analysis for the expressions of MT2 in nonparenchymal cells (NPC) after siRNA-MT2/siRNA-NC was injected into mice tail vein. ( C ) Serum ALT and AST levels were measured. ( D ) Detection of TNF-α, IL1β, IL6, iNOS, and IL10 in the ischemic liver of mice by qRT-PCR. ( E ) HE DCFH, TUNEL, F4/80, CD11b, and Ly6G staining, and ( F, H ) their quantitations. ( G ) Expressions of Bcl-2, Bcl-xl, C-caspase 3, P-NF-κBp65, and NF-κBp65 in IR liver tissues based on Western blotting analysis. ( I ) Detection of TNF-α, IL1β, IL6, iNOS, and IL10 in H/R cocultured XBP1 M-KO BMMs through qRT-PCR. ( J ) Western blotting analysis for the expressions of MT2 in BMMs after transfected with siRNA-MT2/siRNA-NC. ( K ) Migratory abilities of XBP1 M-KO BMMs cocultured for 24 hours based on the Transwell assay. ( L ) Expressions of NLRP3, MT2, IKBα, P-NF-κBp65, and NF-κBp65 in H/R cocultured XBP1 M-KO BMMs using Western blotting assay. n = 6, ∗ P < .05 by Student t test.

Article Snippet: The Suzuki score, which consists of 3 aspects (hepatocyte necrosis, sinusoidal congestion, and ballooning degeneration), was rated on a scale of 1 to 4, as defined by Suzuki et al. Hepatic macrophages and neutrophils were analyzed by IHC staining and incubation with primary rat anti-mouse F4/80, CD11b and Ly6G mAbs (Cell Signaling Technology).

Techniques: Inhibition, In Vivo, In Vitro, Injection, Western Blot, Quantitative RT-PCR, TUNEL Assay, Staining, Transfection, Transwell Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: XBP1 Facilitating NF-κB-p65 Nuclear Translocation Promotes Macrophage-Originated Sterile Inflammation Via Regulating MT2 Transcription in the Ischemia/Reperfusion Liver

doi: 10.1016/j.jcmgh.2024.101402

Figure Lengend Snippet: Sensitizing the NF-kB pathway neutralizes XBP1 M-KO -related benefits in vivo and in vitro. The IRI model mice were subjected to 90-minute ischemia, followed by 6-hour reperfusion. ( A ) XBP1 M-KO mice were treated with BA (20 mg/kg) before establishing hepatic IRI model. ( B ) Serum ALT and AST levels were measured. ( C ) Detection of TNF-α, IL1β, IL6, iNOS, and IL10 in the ischemic liver of mice by qRT-PCR. ( D ) Expressions of Bcl-2, Bcl-xl, C-caspase 3, P-NF-κBp65, and NF-κBp65 in IR liver tissues based on Western blotting analysis. ( E ) HE, DCFH, TUNEL, F4/80, CD11b, and Ly6G staining. ( F ) Western blotting analysis for the expressions of P-NF-κBP65, NF-κBP65 in BMMs after incubated with BA. ( G ) Migratory abilities of BMMs cocultured for 24 hours based on the Transwell assay. ( H ) Expressions of NLRP3, P-NF-κBp65, and NF-κBp65 in H/R cocultured XBP1 M-KO BMMs using Western blotting assay. ( I ) Immunofluorescence staining of MT2 ( red ), NF-κBp65 (purple), and DAPI ( blue ) in H/R cocultured XBP1 M-KO BMMs. ( J ) Detection of TNF-α, IL1β, IL6, iNOS, and IL10 in H/R cocultured XBP1 M-KO BMMs through qRT-PCR. n = 6, ∗ P < .05 by Student t test.

Article Snippet: The Suzuki score, which consists of 3 aspects (hepatocyte necrosis, sinusoidal congestion, and ballooning degeneration), was rated on a scale of 1 to 4, as defined by Suzuki et al. Hepatic macrophages and neutrophils were analyzed by IHC staining and incubation with primary rat anti-mouse F4/80, CD11b and Ly6G mAbs (Cell Signaling Technology).

Techniques: In Vivo, In Vitro, Quantitative RT-PCR, Western Blot, TUNEL Assay, Staining, Incubation, Transwell Assay, Immunofluorescence