bax primary antibodies  (Proteintech)

Journal: Acta Pharmaceutica Sinica. B

Article Title: Natural killer cell-derived granzyme B as a therapeutic target for alleviating graft injury during liver transplantation

doi: 10.1016/j.apsb.2025.07.042

Figure Lengend Snippet: Systemic NK depletion attenuates hepatic IRI in vivo . (A) Schematic image of experiments ( n = 6). (B) Body weight record of the mouse receiving NK1.1 mAb or control a week before hepatic IRI. (C) Flow cytometry analysis of NK cells among total CD45 + cells in liver and blood, respectively. (D) Flow cytometry analysis of total GZMB + cells among total CD45 + cells in the liver. (E) Intra-hepatic tissue level of GZMB measured by ELISA and relatively quantified by total protein concentration. (F) Representative immunofluorescence staining of NK1.1 and GZMB in liver (scale bar = 50 μm). (G) Plasma level of ALT and AST. (H) Representative images of HE staining of mouse liver and analysis of Suzuki score (scale bar = 100 μm). (I) Relative mRNA expression of Gzmb , Il-1β , Tnf-α , and Mmp9 to β- actin in mouse liver. (J) Relative protein level of BAX/BCL2 and cleaved caspase3 (C-CAS3) in mouse liver. GZMB, granzyme B; NK, natural killer; NC, negative control. Data are represented as mean ± SEM. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001; ns, not significant.

Article Snippet: The PVDF membranes were blocked with 5% BSA for 1 h at room temperature and incubated overnight at 4 °C with specific primary antibodies against BAX (CST, MA, USA), BCL2 (abcam, Cambridge, UK), C-CAS3 (CST, MA, USA), β -actin (ABclonal, Wuhan, China).

Techniques: In Vivo, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Protein Concentration, Immunofluorescence, Staining, Clinical Proteomics, Expressing, Negative Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Natural killer cell-derived granzyme B as a therapeutic target for alleviating graft injury during liver transplantation

doi: 10.1016/j.apsb.2025.07.042

Figure Lengend Snippet: NK cells trigger hepatocyte apoptosis and deteriorate liver function through the production of GZMB. (A) Experimental process of NK cells adoptive transfer and murine hepatic IRI model. (B) Absolute quantification of NK cells in spleens and livers after hepatic IRI from immunodeficient or adoptively transferred mice (NC, n = 3; WT-NK, n = 5; KO-NK, n = 6). (C) Representative images of HE staining (scale bar = 200 μm, dashed lines delineate the necrotic areas) and IHC for NK1.1, GZMB, and cleaved caspase3 (C-CAS3) (NC, n = 5; WT-NK, n = 5; KO-NK, n = 6) (scale bar = 100 μm). (D) Mouse plasma level of ALT and AST (Sham, n = 5; IRI, n = 7; NC, n = 5; WT-NK, n = 5; KO-NK, n = 6). (E) Relative mRNA expression of Il-1β , Tnf-α and Mmp9 to β -actin in mouse liver (Sham, n = 5; IRI, n = 7; NC, n = 5; WT-NK, n = 5; KO-NK, n = 6). (F) Relative protein level of BAX, BCL2, and C-CAS3 measured by Western blot ( n = 3). GZMB, granzyme B; NK, natural killer; IRI, ischemia reperfusion injury; NC, negative control; WT, wildtype; KO, knockout; Il-1β, interleukin-1 β ; Tnf-α , tumor necrosis factor α ; Mmp9 , metalloproteinase 9. Data are represented as mean ± SEM. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001; ns, not significant.

Article Snippet: The PVDF membranes were blocked with 5% BSA for 1 h at room temperature and incubated overnight at 4 °C with specific primary antibodies against BAX (CST, MA, USA), BCL2 (abcam, Cambridge, UK), C-CAS3 (CST, MA, USA), β -actin (ABclonal, Wuhan, China).

Techniques: Adoptive Transfer Assay, Quantitative Proteomics, Staining, Clinical Proteomics, Expressing, Western Blot, Negative Control, Knock-Out

Journal: Acta Pharmaceutica Sinica. B

Article Title: Natural killer cell-derived granzyme B as a therapeutic target for alleviating graft injury during liver transplantation

doi: 10.1016/j.apsb.2025.07.042

Figure Lengend Snippet: AEBSF inhibits GZMB and improves liver function in both murine hepatic IRI and rat LT model (A) Schematic image of the experiments ( n = 6). (B) Mouse serum levels of ALT and AST. (C) Relative quantification of GZMB in serum and liver by ELISA. (D) Representative images of HE staining (scale bar = 100 μm, dashed lines delineate the necrotic areas) and IHC for GZMB (scale bar = 100 μm) in mouse liver. (E) Relative gene expression of Il-1β , Tnf-α , and Mmp9 to β-actin in mouse liver tissues. (F) Representative images of immunofluorescence staining of TUNEL in mouse liver (scale bar = 100 μm). (G) Relative protein level of BAX, BCL2, and cleaved caspase3 (C-CAS3) measured by Western blot. (H) Schematic image of rat LT experimental process ( n = 6). (I) Rat serum levels of ALT and AST. (J) Representative images of HE staining (scale bar = 200 μm, dashed lines delineate the necrotic areas), and IHC for GZMB and C-CAS3 (scale bar = 100 μm) in rat livers. LT, liver transplantation; GZMB, granzyme B; C-CAS3, cleaved caspase3. Data are represented as mean ± SEM. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ns, not significant.

Article Snippet: The PVDF membranes were blocked with 5% BSA for 1 h at room temperature and incubated overnight at 4 °C with specific primary antibodies against BAX (CST, MA, USA), BCL2 (abcam, Cambridge, UK), C-CAS3 (CST, MA, USA), β -actin (ABclonal, Wuhan, China).

Techniques: Quantitative Proteomics, Enzyme-linked Immunosorbent Assay, Staining, Gene Expression, Immunofluorescence, TUNEL Assay, Western Blot, Transplantation Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Natural killer cell-derived granzyme B as a therapeutic target for alleviating graft injury during liver transplantation

doi: 10.1016/j.apsb.2025.07.042

Figure Lengend Snippet: FDA-approved vidarabine phosphate functions as a potent GZMB inhibitor to reduce hepatic IRI. (A) 3D structure of mouse GZMB protein and small molecule Vida, wherein the active site Ser203 inside the catalytic pocket of mouse GZMB and Vida are both in stick representations. (B) Time-dependent distance between the phosphate group of Vida and the hydroxyl group of Ser203. Inset: An MD simulation snapshot of Vida in complex with mouse GZMB. (C) The contact potential surface of mouse GZMB, blue (red), represents positive (negative) contact potential. (D) Time evolution of interaction energy between vidarabine phosphate and mouse GZMB. In all the stick representations in (A–C), yellow indicates phosphorus atoms, red oxygen atoms, green carbon atoms, blue nitrogen atoms, and white hydrogen atoms. (E) Schematic image of the experiments (Sham, Cyta, n = 5; NC, Vida, n = 6). (F) Quantification of GZMB in mouse serum and liver by ELISA. (G) Intra-hepatic tissue level of GZMB measured by ELISA and relatively quantified by total protein concentration. (H) Representative images of IHC for GZMB and cleaved caspase3 (C-CAS3) (scale bar = 100 μm) and HE staining (scale bar = 100 μm, dashed lines delineate the necrotic areas). (I) Mouse serum levels of ALT and AST. (J) Relative mRNA expression of Il-1β , Tnf-α , and Mmp9 to β- actin in mouse liver. (K) Relative protein level of BAX, BCL2, and C-CAS3 measured by Western blot ( n = 3). GZMB, granzyme B; NK, natural killer; IRI, ischemia reperfusion injury; NC, negative control; Vida, vidarabine phosphate; Cyta, cytarabine; Il-1β , interleukin-1 β ; Tnf-α , tumor necrosis factor α ; Mmp9 , metalloproteinase 9. Data are represented as mean ± SEM. ∗ P < 0.05; ∗∗ P < 0.01; ns, not significant.

Article Snippet: The PVDF membranes were blocked with 5% BSA for 1 h at room temperature and incubated overnight at 4 °C with specific primary antibodies against BAX (CST, MA, USA), BCL2 (abcam, Cambridge, UK), C-CAS3 (CST, MA, USA), β -actin (ABclonal, Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Protein Concentration, Staining, Expressing, Western Blot, Negative Control