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aspp2 mouse polyclonal antibodies  (Danaher Inc)


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    Danaher Inc aspp2 mouse polyclonal antibodies
    <t>ASPP2</t> is increased in the livers of ALI patients and mouse models induced by D-GalN/LPS. (A–C) Liver tissue obtained from normal controls, patients with CHB and HBV-ALI were analyzed the expression level of ASPP2. (A) Gene expressions of ASPP2 was measured by qRT-PCR. (B) Protein expression levels of ASPP2 was measured by western blot assays. (C) The expression of ASPP2 was measured by immunofluorescence staining. (D–F) Mice were intraperitoneally injected with D-GalN (700 mg/kg) and LPS (10 μg/kg) at 2-, 4- and 6- hours (12 mice/group). The mice in the control group (n = 8) were injected with PBS only. (D) Serum AST and ALT enzyme levels. (E) Gene expressions of ASPP2 was measured by qRT-PCR. (F) Protein expression levels of ASPP2 was measured by western blot assays. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.
    Aspp2 Mouse Polyclonal Antibodies, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/aspp2+mouse+polyclonal+antibodies/pmc11150554-88-3-8
    Average 86 stars, based on 1 article reviews
    aspp2 mouse polyclonal antibodies - by Bioz Stars, 2026-09
    86/100 stars

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    1) Product Images from "Enhancing ASPP2 promotes acute liver injury via an inflammatory immunoregulatory mechanism"

    Article Title: Enhancing ASPP2 promotes acute liver injury via an inflammatory immunoregulatory mechanism

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2024.1381735

    ASPP2 is increased in the livers of ALI patients and mouse models induced by D-GalN/LPS. (A–C) Liver tissue obtained from normal controls, patients with CHB and HBV-ALI were analyzed the expression level of ASPP2. (A) Gene expressions of ASPP2 was measured by qRT-PCR. (B) Protein expression levels of ASPP2 was measured by western blot assays. (C) The expression of ASPP2 was measured by immunofluorescence staining. (D–F) Mice were intraperitoneally injected with D-GalN (700 mg/kg) and LPS (10 μg/kg) at 2-, 4- and 6- hours (12 mice/group). The mice in the control group (n = 8) were injected with PBS only. (D) Serum AST and ALT enzyme levels. (E) Gene expressions of ASPP2 was measured by qRT-PCR. (F) Protein expression levels of ASPP2 was measured by western blot assays. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.
    Figure Legend Snippet: ASPP2 is increased in the livers of ALI patients and mouse models induced by D-GalN/LPS. (A–C) Liver tissue obtained from normal controls, patients with CHB and HBV-ALI were analyzed the expression level of ASPP2. (A) Gene expressions of ASPP2 was measured by qRT-PCR. (B) Protein expression levels of ASPP2 was measured by western blot assays. (C) The expression of ASPP2 was measured by immunofluorescence staining. (D–F) Mice were intraperitoneally injected with D-GalN (700 mg/kg) and LPS (10 μg/kg) at 2-, 4- and 6- hours (12 mice/group). The mice in the control group (n = 8) were injected with PBS only. (D) Serum AST and ALT enzyme levels. (E) Gene expressions of ASPP2 was measured by qRT-PCR. (F) Protein expression levels of ASPP2 was measured by western blot assays. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, Injection

    Knockdown of ASPP2 protects against D-GalN/LPS-induced acute liver injury. ASPP2 +/+ mice and ASPP2 +/- mice were intraperitoneally injected with D-GalN/LPS for 6 hours (12 mice/group). (A) Representative livers and H&E staining of livers from different groups. (B) Serum AST and ALT enzyme levels from different groups. (C) Measurement of cytokine levels in the serum of mice in different groups by the Luminex Milliplex ® MAP Kit (TNF-α, IL-6, IL-1β, IL-18, IL-12, IL-10, CXCL-10, IL-13, IL-22). (D) Analysis of the gene expression levels of cytokines (TNF-α, IL-6, IL-1β, IL-18, IL-10, IL-12, CXCL-10) in liver tissues from different groups by qRT-PCR. (E) TUNEL staining (red) liver tissue at 6 h after D-GalN/LPS administration. Original magnification×200. (F) Caspase 3 activity of livers was measured from different groups. (G) Protein expression levels of p-JNK, caspase 3 and cleaved caspase 3 was measured by western blot assays. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.
    Figure Legend Snippet: Knockdown of ASPP2 protects against D-GalN/LPS-induced acute liver injury. ASPP2 +/+ mice and ASPP2 +/- mice were intraperitoneally injected with D-GalN/LPS for 6 hours (12 mice/group). (A) Representative livers and H&E staining of livers from different groups. (B) Serum AST and ALT enzyme levels from different groups. (C) Measurement of cytokine levels in the serum of mice in different groups by the Luminex Milliplex ® MAP Kit (TNF-α, IL-6, IL-1β, IL-18, IL-12, IL-10, CXCL-10, IL-13, IL-22). (D) Analysis of the gene expression levels of cytokines (TNF-α, IL-6, IL-1β, IL-18, IL-10, IL-12, CXCL-10) in liver tissues from different groups by qRT-PCR. (E) TUNEL staining (red) liver tissue at 6 h after D-GalN/LPS administration. Original magnification×200. (F) Caspase 3 activity of livers was measured from different groups. (G) Protein expression levels of p-JNK, caspase 3 and cleaved caspase 3 was measured by western blot assays. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Techniques Used: Injection, Staining, Luminex, Expressing, Quantitative RT-PCR, TUNEL Assay, Activity Assay, Western Blot

    Knockdown of ASPP2 reduced hepatocyte apoptosis through autophagy activation during ALI. (A, B) ASPP2 +/+ mice and ASPP2 +/- mice were intraperitoneally injected with D-GalN/LPS for 6 hours (12 mice/group). (A) Autophagosomes was observed by electron microscope in the livers from different groups. (B) Protein expression levels of LC3, p62, ATG-5, Atg-7 and Beclin-1 were measured by western blotting. (C–E) Mice were pretreated with Atg7 or negative control siRNA (3mg/kg, iv) for 48 hours prior to D-GalN/LPS exposure (n = 12). (C) Serum AST and ALT enzyme levels from different groups. (D) TUNEL staining (red) liver tissue from different groups. Original magnification×200. (E) Caspase 3 activity of livers was measured from different groups. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.
    Figure Legend Snippet: Knockdown of ASPP2 reduced hepatocyte apoptosis through autophagy activation during ALI. (A, B) ASPP2 +/+ mice and ASPP2 +/- mice were intraperitoneally injected with D-GalN/LPS for 6 hours (12 mice/group). (A) Autophagosomes was observed by electron microscope in the livers from different groups. (B) Protein expression levels of LC3, p62, ATG-5, Atg-7 and Beclin-1 were measured by western blotting. (C–E) Mice were pretreated with Atg7 or negative control siRNA (3mg/kg, iv) for 48 hours prior to D-GalN/LPS exposure (n = 12). (C) Serum AST and ALT enzyme levels from different groups. (D) TUNEL staining (red) liver tissue from different groups. Original magnification×200. (E) Caspase 3 activity of livers was measured from different groups. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Techniques Used: Activation Assay, Injection, Microscopy, Expressing, Western Blot, Negative Control, TUNEL Assay, Staining, Activity Assay

    ASPP2-depleted BMMs protect hepatocytes from apoptosis by reducing TNF-α levels in vitro. (A, B) Primary hepatocytes were treated with conditioned medium (CM) of ASPP2 +/+ BMMs or ASPP2 +/- BMMs that with or without LPS (20ng/ml). (A, B) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (C) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. (D) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of TNF-α was analyzed by qRT-PCR. (E) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) as the indicated time points, the level of TNF-α was analyzed by qRT-PCR. (F) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of TNF-α in medium was analyzed by ELISA. *p<0.1, ***p<0.001, ****p<0.0001.
    Figure Legend Snippet: ASPP2-depleted BMMs protect hepatocytes from apoptosis by reducing TNF-α levels in vitro. (A, B) Primary hepatocytes were treated with conditioned medium (CM) of ASPP2 +/+ BMMs or ASPP2 +/- BMMs that with or without LPS (20ng/ml). (A, B) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (C) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. (D) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of TNF-α was analyzed by qRT-PCR. (E) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) as the indicated time points, the level of TNF-α was analyzed by qRT-PCR. (F) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of TNF-α in medium was analyzed by ELISA. *p<0.1, ***p<0.001, ****p<0.0001.

    Techniques Used: In Vitro, CCK-8 Assay, Flow Cytometry, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    ASPP2-depleted BMMs decreased TNF-α by up-regulation of IL-6. (A) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of IL-6 was analyzed by qRT-PCR. E. ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) as the indicated time points, the level of IL-6 was analyzed by qRT-PCR. E. ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of IL-6 in medium was analyzed by ELISA. (D) IL-6 siRNA or negative control siRNA was transected into ASPP2 +/+ BMMs and ASPP2 +/- BMMs that with or without LPS (20ng/ml) stimulation. The level of TNF-α in BMMs were analyzed by qRT-PCR. (E, F) primary hepatocytes were treated with conditioned medium (CM) collected from (D) . The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (G) Primary hepatocytes were treated with CM the same as (D) . The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.
    Figure Legend Snippet: ASPP2-depleted BMMs decreased TNF-α by up-regulation of IL-6. (A) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of IL-6 was analyzed by qRT-PCR. E. ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) as the indicated time points, the level of IL-6 was analyzed by qRT-PCR. E. ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of IL-6 in medium was analyzed by ELISA. (D) IL-6 siRNA or negative control siRNA was transected into ASPP2 +/+ BMMs and ASPP2 +/- BMMs that with or without LPS (20ng/ml) stimulation. The level of TNF-α in BMMs were analyzed by qRT-PCR. (E, F) primary hepatocytes were treated with conditioned medium (CM) collected from (D) . The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (G) Primary hepatocytes were treated with CM the same as (D) . The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Techniques Used: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Negative Control, CCK-8 Assay, Flow Cytometry

    ASPP2 +/− BMMs promote IL-6 expression through autophagy induction. (A, B) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 12 hours. (A) The gene expression levels of autophagy related genes (LC3, ATG-7, ATG-5, Beclin-1) were analyzed by qRT-PCR. (B) The protein expression levels of autophagy related genes (LC3, ATG-7, ATG-5, Beclin-1) were analyzed by western blotting. (C) DMSO and 3-MA were pretreated with cells 2 hours before LPS stimulation. The gene expression levels of cytokines (IL-6, TNF-α) in cells were analyzed by qRT-PCR. (D, E) Primary hepatocytes were incubated with conditioned medium (CM) of BMMs treated as (C) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (F) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. *p<0.1, **p<0.01, ****p<0.0001.
    Figure Legend Snippet: ASPP2 +/− BMMs promote IL-6 expression through autophagy induction. (A, B) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 12 hours. (A) The gene expression levels of autophagy related genes (LC3, ATG-7, ATG-5, Beclin-1) were analyzed by qRT-PCR. (B) The protein expression levels of autophagy related genes (LC3, ATG-7, ATG-5, Beclin-1) were analyzed by western blotting. (C) DMSO and 3-MA were pretreated with cells 2 hours before LPS stimulation. The gene expression levels of cytokines (IL-6, TNF-α) in cells were analyzed by qRT-PCR. (D, E) Primary hepatocytes were incubated with conditioned medium (CM) of BMMs treated as (C) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (F) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. *p<0.1, **p<0.01, ****p<0.0001.

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Incubation, CCK-8 Assay, Flow Cytometry

    Upregulation of ASPP2 sensitizes hepatocytes to TNF-α-induced apoptosis. (A-C) Primary hepatocytes were transfected with ASPP2 over-expression (OE) plasmid or ASPP2 siRNA, and treated with TNF-α for 12 hours. (A, B) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (C) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. (D) The graphic abstract of this study. In ALI mice induced by D-GalN/LPS, ASPP2 was increased in liver Kupffer cells, which suppressed the level of IL-6 and increased the level of TNF-α through inhibition autophagy to promote hepatocytes apoptosis. In addition, upregulation of ASPP2 in hepatocytes promoted cell apoptosis induced by TNF-α. *p<0.1, ***p<0.001, ****p<0.0001.
    Figure Legend Snippet: Upregulation of ASPP2 sensitizes hepatocytes to TNF-α-induced apoptosis. (A-C) Primary hepatocytes were transfected with ASPP2 over-expression (OE) plasmid or ASPP2 siRNA, and treated with TNF-α for 12 hours. (A, B) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (C) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. (D) The graphic abstract of this study. In ALI mice induced by D-GalN/LPS, ASPP2 was increased in liver Kupffer cells, which suppressed the level of IL-6 and increased the level of TNF-α through inhibition autophagy to promote hepatocytes apoptosis. In addition, upregulation of ASPP2 in hepatocytes promoted cell apoptosis induced by TNF-α. *p<0.1, ***p<0.001, ****p<0.0001.

    Techniques Used: Transfection, Over Expression, Plasmid Preparation, CCK-8 Assay, Flow Cytometry, Inhibition

    Related Articles

    Incubation:

    Article Title: Enhancing ASPP2 promotes acute liver injury via an inflammatory immunoregulatory mechanism
    Article Snippet: The slides were permeabilized in 0.01% Triton X-100, and then blocked in 10% goat serum in 3% bovine serum albumin (BSA, Sigma, St. Louis, MO, USA) for 60 minutes. .. Then, incubated with ASPP2 mouse polyclonal antibodies (1:200, Abcam, Cambridge, MA) diluted in 3% BSA overnight at 4°C. .. After washing with PBS three times, the slides were incubated with Alexa Fluor ® 568 goat anti-rabbit IgG (1:200, Invitrogen, Grand Island, NY) for 45 minutes at room temperature.



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    aspp2 mouse polyclonal antibodies  (Danaher Inc)
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    Danaher Inc aspp2 mouse polyclonal antibodies
    <t>ASPP2</t> is increased in the livers of ALI patients and mouse models induced by D-GalN/LPS. (A–C) Liver tissue obtained from normal controls, patients with CHB and HBV-ALI were analyzed the expression level of ASPP2. (A) Gene expressions of ASPP2 was measured by qRT-PCR. (B) Protein expression levels of ASPP2 was measured by western blot assays. (C) The expression of ASPP2 was measured by immunofluorescence staining. (D–F) Mice were intraperitoneally injected with D-GalN (700 mg/kg) and LPS (10 μg/kg) at 2-, 4- and 6- hours (12 mice/group). The mice in the control group (n = 8) were injected with PBS only. (D) Serum AST and ALT enzyme levels. (E) Gene expressions of ASPP2 was measured by qRT-PCR. (F) Protein expression levels of ASPP2 was measured by western blot assays. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.
    Aspp2 Mouse Polyclonal Antibodies, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/aspp2+mouse+polyclonal+antibodies/pmc11150554-88-3-8
    Average 86 stars, based on 1 article reviews
    aspp2 mouse polyclonal antibodies - by Bioz Stars, 2026-09
    86/100 stars
    		  Buy from Supplier

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    ASPP2 is increased in the livers of ALI patients and mouse models induced by D-GalN/LPS. (A–C) Liver tissue obtained from normal controls, patients with CHB and HBV-ALI were analyzed the expression level of ASPP2. (A) Gene expressions of ASPP2 was measured by qRT-PCR. (B) Protein expression levels of ASPP2 was measured by western blot assays. (C) The expression of ASPP2 was measured by immunofluorescence staining. (D–F) Mice were intraperitoneally injected with D-GalN (700 mg/kg) and LPS (10 μg/kg) at 2-, 4- and 6- hours (12 mice/group). The mice in the control group (n = 8) were injected with PBS only. (D) Serum AST and ALT enzyme levels. (E) Gene expressions of ASPP2 was measured by qRT-PCR. (F) Protein expression levels of ASPP2 was measured by western blot assays. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Journal: Frontiers in Immunology

    Article Title: Enhancing ASPP2 promotes acute liver injury via an inflammatory immunoregulatory mechanism

    doi: 10.3389/fimmu.2024.1381735

    Figure Lengend Snippet: ASPP2 is increased in the livers of ALI patients and mouse models induced by D-GalN/LPS. (A–C) Liver tissue obtained from normal controls, patients with CHB and HBV-ALI were analyzed the expression level of ASPP2. (A) Gene expressions of ASPP2 was measured by qRT-PCR. (B) Protein expression levels of ASPP2 was measured by western blot assays. (C) The expression of ASPP2 was measured by immunofluorescence staining. (D–F) Mice were intraperitoneally injected with D-GalN (700 mg/kg) and LPS (10 μg/kg) at 2-, 4- and 6- hours (12 mice/group). The mice in the control group (n = 8) were injected with PBS only. (D) Serum AST and ALT enzyme levels. (E) Gene expressions of ASPP2 was measured by qRT-PCR. (F) Protein expression levels of ASPP2 was measured by western blot assays. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Article Snippet: Then, incubated with ASPP2 mouse polyclonal antibodies (1:200, Abcam, Cambridge, MA) diluted in 3% BSA overnight at 4°C.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, Injection

    Knockdown of ASPP2 protects against D-GalN/LPS-induced acute liver injury. ASPP2 +/+ mice and ASPP2 +/- mice were intraperitoneally injected with D-GalN/LPS for 6 hours (12 mice/group). (A) Representative livers and H&E staining of livers from different groups. (B) Serum AST and ALT enzyme levels from different groups. (C) Measurement of cytokine levels in the serum of mice in different groups by the Luminex Milliplex ® MAP Kit (TNF-α, IL-6, IL-1β, IL-18, IL-12, IL-10, CXCL-10, IL-13, IL-22). (D) Analysis of the gene expression levels of cytokines (TNF-α, IL-6, IL-1β, IL-18, IL-10, IL-12, CXCL-10) in liver tissues from different groups by qRT-PCR. (E) TUNEL staining (red) liver tissue at 6 h after D-GalN/LPS administration. Original magnification×200. (F) Caspase 3 activity of livers was measured from different groups. (G) Protein expression levels of p-JNK, caspase 3 and cleaved caspase 3 was measured by western blot assays. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Journal: Frontiers in Immunology

    Article Title: Enhancing ASPP2 promotes acute liver injury via an inflammatory immunoregulatory mechanism

    doi: 10.3389/fimmu.2024.1381735

    Figure Lengend Snippet: Knockdown of ASPP2 protects against D-GalN/LPS-induced acute liver injury. ASPP2 +/+ mice and ASPP2 +/- mice were intraperitoneally injected with D-GalN/LPS for 6 hours (12 mice/group). (A) Representative livers and H&E staining of livers from different groups. (B) Serum AST and ALT enzyme levels from different groups. (C) Measurement of cytokine levels in the serum of mice in different groups by the Luminex Milliplex ® MAP Kit (TNF-α, IL-6, IL-1β, IL-18, IL-12, IL-10, CXCL-10, IL-13, IL-22). (D) Analysis of the gene expression levels of cytokines (TNF-α, IL-6, IL-1β, IL-18, IL-10, IL-12, CXCL-10) in liver tissues from different groups by qRT-PCR. (E) TUNEL staining (red) liver tissue at 6 h after D-GalN/LPS administration. Original magnification×200. (F) Caspase 3 activity of livers was measured from different groups. (G) Protein expression levels of p-JNK, caspase 3 and cleaved caspase 3 was measured by western blot assays. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Article Snippet: Then, incubated with ASPP2 mouse polyclonal antibodies (1:200, Abcam, Cambridge, MA) diluted in 3% BSA overnight at 4°C.

    Techniques: Injection, Staining, Luminex, Expressing, Quantitative RT-PCR, TUNEL Assay, Activity Assay, Western Blot

    Knockdown of ASPP2 reduced hepatocyte apoptosis through autophagy activation during ALI. (A, B) ASPP2 +/+ mice and ASPP2 +/- mice were intraperitoneally injected with D-GalN/LPS for 6 hours (12 mice/group). (A) Autophagosomes was observed by electron microscope in the livers from different groups. (B) Protein expression levels of LC3, p62, ATG-5, Atg-7 and Beclin-1 were measured by western blotting. (C–E) Mice were pretreated with Atg7 or negative control siRNA (3mg/kg, iv) for 48 hours prior to D-GalN/LPS exposure (n = 12). (C) Serum AST and ALT enzyme levels from different groups. (D) TUNEL staining (red) liver tissue from different groups. Original magnification×200. (E) Caspase 3 activity of livers was measured from different groups. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Journal: Frontiers in Immunology

    Article Title: Enhancing ASPP2 promotes acute liver injury via an inflammatory immunoregulatory mechanism

    doi: 10.3389/fimmu.2024.1381735

    Figure Lengend Snippet: Knockdown of ASPP2 reduced hepatocyte apoptosis through autophagy activation during ALI. (A, B) ASPP2 +/+ mice and ASPP2 +/- mice were intraperitoneally injected with D-GalN/LPS for 6 hours (12 mice/group). (A) Autophagosomes was observed by electron microscope in the livers from different groups. (B) Protein expression levels of LC3, p62, ATG-5, Atg-7 and Beclin-1 were measured by western blotting. (C–E) Mice were pretreated with Atg7 or negative control siRNA (3mg/kg, iv) for 48 hours prior to D-GalN/LPS exposure (n = 12). (C) Serum AST and ALT enzyme levels from different groups. (D) TUNEL staining (red) liver tissue from different groups. Original magnification×200. (E) Caspase 3 activity of livers was measured from different groups. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Article Snippet: Then, incubated with ASPP2 mouse polyclonal antibodies (1:200, Abcam, Cambridge, MA) diluted in 3% BSA overnight at 4°C.

    Techniques: Activation Assay, Injection, Microscopy, Expressing, Western Blot, Negative Control, TUNEL Assay, Staining, Activity Assay

    ASPP2-depleted BMMs protect hepatocytes from apoptosis by reducing TNF-α levels in vitro. (A, B) Primary hepatocytes were treated with conditioned medium (CM) of ASPP2 +/+ BMMs or ASPP2 +/- BMMs that with or without LPS (20ng/ml). (A, B) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (C) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. (D) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of TNF-α was analyzed by qRT-PCR. (E) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) as the indicated time points, the level of TNF-α was analyzed by qRT-PCR. (F) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of TNF-α in medium was analyzed by ELISA. *p<0.1, ***p<0.001, ****p<0.0001.

    Journal: Frontiers in Immunology

    Article Title: Enhancing ASPP2 promotes acute liver injury via an inflammatory immunoregulatory mechanism

    doi: 10.3389/fimmu.2024.1381735

    Figure Lengend Snippet: ASPP2-depleted BMMs protect hepatocytes from apoptosis by reducing TNF-α levels in vitro. (A, B) Primary hepatocytes were treated with conditioned medium (CM) of ASPP2 +/+ BMMs or ASPP2 +/- BMMs that with or without LPS (20ng/ml). (A, B) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (C) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. (D) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of TNF-α was analyzed by qRT-PCR. (E) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) as the indicated time points, the level of TNF-α was analyzed by qRT-PCR. (F) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of TNF-α in medium was analyzed by ELISA. *p<0.1, ***p<0.001, ****p<0.0001.

    Article Snippet: Then, incubated with ASPP2 mouse polyclonal antibodies (1:200, Abcam, Cambridge, MA) diluted in 3% BSA overnight at 4°C.

    Techniques: In Vitro, CCK-8 Assay, Flow Cytometry, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    ASPP2-depleted BMMs decreased TNF-α by up-regulation of IL-6. (A) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of IL-6 was analyzed by qRT-PCR. E. ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) as the indicated time points, the level of IL-6 was analyzed by qRT-PCR. E. ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of IL-6 in medium was analyzed by ELISA. (D) IL-6 siRNA or negative control siRNA was transected into ASPP2 +/+ BMMs and ASPP2 +/- BMMs that with or without LPS (20ng/ml) stimulation. The level of TNF-α in BMMs were analyzed by qRT-PCR. (E, F) primary hepatocytes were treated with conditioned medium (CM) collected from (D) . The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (G) Primary hepatocytes were treated with CM the same as (D) . The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Journal: Frontiers in Immunology

    Article Title: Enhancing ASPP2 promotes acute liver injury via an inflammatory immunoregulatory mechanism

    doi: 10.3389/fimmu.2024.1381735

    Figure Lengend Snippet: ASPP2-depleted BMMs decreased TNF-α by up-regulation of IL-6. (A) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of IL-6 was analyzed by qRT-PCR. E. ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) as the indicated time points, the level of IL-6 was analyzed by qRT-PCR. E. ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 6 hours, the level of IL-6 in medium was analyzed by ELISA. (D) IL-6 siRNA or negative control siRNA was transected into ASPP2 +/+ BMMs and ASPP2 +/- BMMs that with or without LPS (20ng/ml) stimulation. The level of TNF-α in BMMs were analyzed by qRT-PCR. (E, F) primary hepatocytes were treated with conditioned medium (CM) collected from (D) . The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (G) Primary hepatocytes were treated with CM the same as (D) . The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. *p<0.1, **p<0.01, ***p<0.001, ****p<0.0001.

    Article Snippet: Then, incubated with ASPP2 mouse polyclonal antibodies (1:200, Abcam, Cambridge, MA) diluted in 3% BSA overnight at 4°C.

    Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Negative Control, CCK-8 Assay, Flow Cytometry

    ASPP2 +/− BMMs promote IL-6 expression through autophagy induction. (A, B) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 12 hours. (A) The gene expression levels of autophagy related genes (LC3, ATG-7, ATG-5, Beclin-1) were analyzed by qRT-PCR. (B) The protein expression levels of autophagy related genes (LC3, ATG-7, ATG-5, Beclin-1) were analyzed by western blotting. (C) DMSO and 3-MA were pretreated with cells 2 hours before LPS stimulation. The gene expression levels of cytokines (IL-6, TNF-α) in cells were analyzed by qRT-PCR. (D, E) Primary hepatocytes were incubated with conditioned medium (CM) of BMMs treated as (C) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (F) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. *p<0.1, **p<0.01, ****p<0.0001.

    Journal: Frontiers in Immunology

    Article Title: Enhancing ASPP2 promotes acute liver injury via an inflammatory immunoregulatory mechanism

    doi: 10.3389/fimmu.2024.1381735

    Figure Lengend Snippet: ASPP2 +/− BMMs promote IL-6 expression through autophagy induction. (A, B) ASPP2 +/+ BMMs or ASPP2 +/- BMMs were treated with LPS (20ng/ml) for 12 hours. (A) The gene expression levels of autophagy related genes (LC3, ATG-7, ATG-5, Beclin-1) were analyzed by qRT-PCR. (B) The protein expression levels of autophagy related genes (LC3, ATG-7, ATG-5, Beclin-1) were analyzed by western blotting. (C) DMSO and 3-MA were pretreated with cells 2 hours before LPS stimulation. The gene expression levels of cytokines (IL-6, TNF-α) in cells were analyzed by qRT-PCR. (D, E) Primary hepatocytes were incubated with conditioned medium (CM) of BMMs treated as (C) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (F) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. *p<0.1, **p<0.01, ****p<0.0001.

    Article Snippet: Then, incubated with ASPP2 mouse polyclonal antibodies (1:200, Abcam, Cambridge, MA) diluted in 3% BSA overnight at 4°C.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Incubation, CCK-8 Assay, Flow Cytometry

    Upregulation of ASPP2 sensitizes hepatocytes to TNF-α-induced apoptosis. (A-C) Primary hepatocytes were transfected with ASPP2 over-expression (OE) plasmid or ASPP2 siRNA, and treated with TNF-α for 12 hours. (A, B) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (C) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. (D) The graphic abstract of this study. In ALI mice induced by D-GalN/LPS, ASPP2 was increased in liver Kupffer cells, which suppressed the level of IL-6 and increased the level of TNF-α through inhibition autophagy to promote hepatocytes apoptosis. In addition, upregulation of ASPP2 in hepatocytes promoted cell apoptosis induced by TNF-α. *p<0.1, ***p<0.001, ****p<0.0001.

    Journal: Frontiers in Immunology

    Article Title: Enhancing ASPP2 promotes acute liver injury via an inflammatory immunoregulatory mechanism

    doi: 10.3389/fimmu.2024.1381735

    Figure Lengend Snippet: Upregulation of ASPP2 sensitizes hepatocytes to TNF-α-induced apoptosis. (A-C) Primary hepatocytes were transfected with ASPP2 over-expression (OE) plasmid or ASPP2 siRNA, and treated with TNF-α for 12 hours. (A, B) The cell viability and cell death rate of primary hepatocytes were measured by CCK-8 assays and LDH assays, respectively. (C) The cell apoptosis rate of primary hepatocytes was measured by flow cytometry. (D) The graphic abstract of this study. In ALI mice induced by D-GalN/LPS, ASPP2 was increased in liver Kupffer cells, which suppressed the level of IL-6 and increased the level of TNF-α through inhibition autophagy to promote hepatocytes apoptosis. In addition, upregulation of ASPP2 in hepatocytes promoted cell apoptosis induced by TNF-α. *p<0.1, ***p<0.001, ****p<0.0001.

    Article Snippet: Then, incubated with ASPP2 mouse polyclonal antibodies (1:200, Abcam, Cambridge, MA) diluted in 3% BSA overnight at 4°C.

    Techniques: Transfection, Over Expression, Plasmid Preparation, CCK-8 Assay, Flow Cytometry, Inhibition