antibody against ephb2  (Boster Bio)

Journal: Translational Oncology

Article Title: Diagnostic and prognostic value of EphB2 in nasopharyngeal carcinoma

doi: 10.1016/j.tranon.2025.102641

Figure Lengend Snippet: Comparison of serum EphB2 levels between NPC patients and the control group. Data are presented as mean ± SD.

Article Snippet: The membranes were then incubated with a primary antibody against EphB2 (BOSTER, #A01507–1) overnight at 4 °C, followed by incubation with an HRP-conjugated secondary antibody.

Techniques: Comparison, Control

Journal: Translational Oncology

Article Title: Diagnostic and prognostic value of EphB2 in nasopharyngeal carcinoma

doi: 10.1016/j.tranon.2025.102641

Figure Lengend Snippet: EphB2 protein expression in nasopharyngeal tissues. (A) Representative western blot images of EphB2 expression in control tissues (nasopharyngeal mucosal chronic inflammation) and NPC tissues stratified by clinical stage (I–II vs. III–IV), EBV status, and recurrence status ( n = 3 per subgroup). Approximate molecular weights: EphB2, 117 kDa; GAPDH, 36 kDa. (B) Quantitative analysis of EphB2 expression normalized to GAPDH. Data represent mean ± SD from six independent experiments. ** P < 0.01 by one-way ANOVA with Tukey’s post-hoc test.

Article Snippet: The membranes were then incubated with a primary antibody against EphB2 (BOSTER, #A01507–1) overnight at 4 °C, followed by incubation with an HRP-conjugated secondary antibody.

Techniques: Expressing, Western Blot, Control

Journal: Translational Oncology

Article Title: Diagnostic and prognostic value of EphB2 in nasopharyngeal carcinoma

doi: 10.1016/j.tranon.2025.102641

Figure Lengend Snippet: Immunofluorescence analysis of EphB2 expression in nasopharyngeal tissues. A: Nasopharyngeal mucosal chronic inflammation tissues; B: EBV(-) NPC tissues; C: EBV(+) NPC tissues. EphB2 staining is shown in red; nuclei are counterstained with DAPI (blue). Original magnification: 400×.

Article Snippet: The membranes were then incubated with a primary antibody against EphB2 (BOSTER, #A01507–1) overnight at 4 °C, followed by incubation with an HRP-conjugated secondary antibody.

Techniques: Immunofluorescence, Expressing, Staining

Journal: Translational Oncology

Article Title: Diagnostic and prognostic value of EphB2 in nasopharyngeal carcinoma

doi: 10.1016/j.tranon.2025.102641

Figure Lengend Snippet: Disease-free survival analysis based on recurrence (A), EBV infection status (B), and serum EphB2 expression (C). In (C), patients were divided into EphB2-Low and EphB2-High groups using the median serum concentration (11.161 ng/mL) as the cutoff. Vertical ticks indicate censored data.

Article Snippet: The membranes were then incubated with a primary antibody against EphB2 (BOSTER, #A01507–1) overnight at 4 °C, followed by incubation with an HRP-conjugated secondary antibody.

Techniques: Infection, Expressing, Concentration Assay

Journal: Translational Oncology

Article Title: Diagnostic and prognostic value of EphB2 in nasopharyngeal carcinoma

doi: 10.1016/j.tranon.2025.102641

Figure Lengend Snippet: ROC curves evaluating the diagnostic performance of serum EphB2, EBV infection status, and their combination in discriminating NPC patients from controls.

Article Snippet: The membranes were then incubated with a primary antibody against EphB2 (BOSTER, #A01507–1) overnight at 4 °C, followed by incubation with an HRP-conjugated secondary antibody.

Techniques: Diagnostic Assay, Infection

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: ( A ) Analysis of human MASLD/MASH RNA-seq data (GSE130970) showing that EPHB2 transcript is elevated in advanced MASH (* p < 0.05 by ANOVA). ( B ) Representative images of liver sections from human with MASLD/MASH at various stage of fibrosis, stained with H&E and phosphoEphB2. Images were taken using a 20x objective. ( C ) Quantification of phosphoEphB2 in liver specimen of human with MASLD/MASH (n =4-10/group, ** p < 0.01 by ANOVA). ( D ) Analysis of murine MASH RNA-seq data (GSE119340) showing upregulation of liver Ephb2 transcript in MASH (n=3/group, * p <0.05 by Student t test). ( E ) Liver mRNA levels of Ephb2 , Vim , Cd11b in mice fed the obesogenic GAN diet at week 7, 12, 18, and 26 (n = 3-5 mice/group, * p < 0.05, ** p < 0.01, *** p <0.001, **** p <0.0001 by ANOVA). ( F ) Representative images of immunohistochemistry of phosphoEphB2 in liver sections of chow and GAN fed mice at week 26. Images were taken using the 10x objectives. ( G ) Western blots images showing proteins levels of EphB2, αSMA and GAPDH in liver samples of chow and CDAA-HFD fed mice for 12 weeks (n=3 mice/group). Data are presented as mean ± SEM, ns = not significant.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: RNA Sequencing, Staining, Immunohistochemistry, Western Blot

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: ( A ) Body weight gained in Ephb2+/+ and Ephb2-/- mice fed the obesogenic GAN diet for 26 weeks. ( B ) Body weight gained in Ephb2+/+ and Ephb2-/- mice fed a rodent chow for 26 weeks. ( C ) Liver to body weight ratio in Ephb2+/+ and Ephb2-/- mice fed the GAN diet for 26 weeks. ( D ) Glucose tolerance testing performed in Ephb2+/+ and Ephb2-/- mice fed the obesogenic GAN diet at week 22. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01, *** p <0.001, **** p <0.0001 by student t test.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques:

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: ( A ) Representative images of liver sections of Ephb2+/+ and Ephb2-/- mice fed the GAN diet for 26 weeks stained with H&E and Masson Trichrome. ( B ) Plasma levels of liver damage enzymes ALT and AST of Ephb2+/+ and Ephb2-/- mice fed the GAN diet for 26 weeks ( n = 4 mice/group, * p < 0.05, ** p < 0.01 by Student’s t test). ( C ) Plasma level of cholesterol and triglyceride from Ephb2+/+ and Ephb2-/- mice fed the GAN diet for 26 weeks ( n = 4 mice/group, * p < 0.05, ** p < 0.01 by Student’s t test). ( D ) Heatmap depicting the top 40 differentially expressed genes from bulk RNA sequencing of livers samples of Ephb2+/+ and Ephb2-/- mice fed the GAN diet for 26 weeks. ( E ) Gene ontology analysis of the top significantly downregulated or upregulated pathways in RNA sequencing data in D . ( F ) Liver mRNA levels of fibrotic markers Col1a1, Timp1, Edn1, Acta2, Pdgfr β, Vim and ( G ) inflammatory markers, Cd11b, Il-6, Tnf- α, Ccr2, Tgf- β 1 , and Il-1 β in Ephb2+/+ and Ephb2-/- mice fed the GAN diet for 26 weeks ( n = 5 mice/group, * p < 0.05, ** p < 0.01 by Student’s t test). ( H ) Representative images of liver sections of Ephb2+/+ and Ephb2-/- mice fed the CDAA-HFD diet for 12 weeks stained with H&E and Masson Trichrome. ( I ) Plasma levels of liver damage enzymes ALT and AST in Ephb2+/+ and Ephb2-/- mice fed the CDAA-HFD ( n = 9-11 mice/group, * p < 0.05 by Student’s t test). ( J ) Plasma level of cholesterol and triglyceride in Ephb2+/+ and Ephb2-/- mice fed the CDAA-HFD ( n = 9-11 mice/group, * p < 0.05, ** p < 0.01 by Student’s t test. ( K ) Level of hydroxyproline quantified in the livers of Ephb2+/+ and Ephb2-/- mice fed the CDAA-HFD ( n = 7-10 mice per group, * p < 0.05, ** p < 0.01 by Student’s t test). ( L ) Liver mRNA levels of fibrotic and inflammatory markers, Col1a1, Col6a2, Acta2, Tgf- β 1, Smad3, Il-1 β, Il-6, Cd11b in Ephb2+/+ and Ephb2-/- mice fed the CDAA-HFD for 12 weeks ( n = 9-11 mice/group, * p < 0.05, ** p < 0.01 by Student’s t test). Data are presented as mean ± SEM, ns = not significant.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: Staining, Clinical Proteomics, RNA Sequencing

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: ( A ) Gating strategy used to identify various immune cells present in the livers of Ephb2+/+ and Ephb2-/- mice. ( B ) Quantification of hepatic myeloid cells subsets and T (CD3) and B (CD19) cells in the livers of Ephb2+/+ and Ephb2-/- mice fed the obesogenic GAN diet (n = 5 mice/group, * p < 0.05 by student t test). ( C ) Immunophenotyping of macrophages subset (Ly6C, F4/80, PNM-neutrophils) dendritic cells (cDC1) in the livers of Ephb2+/+ and Ephb2 -/- mice fed the non-obesogenic CDAA-HFD for 12 weeks (n=9-10 mice/group, ** p < 0.01 by Student t test). Data are presented as mean ± SEM, ns = not significant.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques:

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: ( A ) Representative images of livers from EphB2-F620D, wild type and EphB2-K661R mice fed the GAN diet for 26 weeks photographed, and sections stained with H&E and Masson Trichrome. ( B ) Level of hydroxyproline quantified in the livers of EphB2-F620D, wild type and EphB2-K661R mice fed the GAN diet for 26 weeks (n = 8 mice/group). ( C ) Plasma levels of liver damage enzymes ALT and AST in EphB2-F620D, wild type and EphB2-K661R mice fed the GAN diet (n=8-11 mice/group, ** p < 0.01 by ANOVA test. ( D ) plasma level of cholesterol and triglycerides ( E ) Liver to body weight ratio of EphB2-F620D, wild type and EphB2-K661R mice fed the GAN diet ( n = 8-11 mice per group, * p < 0.05, ** p < 0.01, *** p < 0.001 by ANOVA). ( F ) Liver mRNA levels of fibrotic and inflammatory markers Col1a1, Col3a1, Col6a2, Tgf- β 1, Il-1 β, Il-6, Cd11b in EphB2-F620D, wild type and EphB2-K661R mice fed the GAN diet for 26 weeks ( n = 8-11 mice/group, * p < 0.05, ** p < 0.01, by ANOVA). ( G ) Representative images of livers section of EphB2-F620D, wild type and EphB2-K661R receiving CCl 4 for 4 weeks and stained with H&E , Masson Trichrome and Sirius red. ( H ) Level of hydroxyproline quantified in the livers of EphB2-F620D, wild type and EphB2-K661R mice receiving CCl 4 for 4 weeks (n = 4-5 mice/group). ( I ) Plasma levels of liver damage enzymes ALT and AST in EphB2-F620D, wild type and EphB2-K661R mice (n= 4-5 mice/group, * p < 0.05, ** p < 0.01 by ANOVA). ( J ) Percentage of myeloid cell subsets population (CD11b hi , cDC1, F4/80) in the liver of EphB2-F620D, wild type and EphB2-K661R mice receiving CCl 4 for 4 weeks (n= 4-5 mice/group, * p < 0.05, ** p < 0.01, **** p < 0.0001 by ANOVA). ( K ) Liver mRNA levels of fibrotic markers Col1a1, Col3a1, Acta2, Timp1, Col1a2 , and Col6a2 in EphB2-F620D, wild type and EphB2-K661R mice receiving CCl 4 for 4 weeks ( n = 4-5 mice/group, * p < 0.05, ** p < 0.01 by ANOVA). Data are presented as mean ± SEM, ns = not significant.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: Staining, Clinical Proteomics

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: ( A ) Representative immunofluorescence images of livers sections from Ephb2 fl/fl (Hep WT ) and Hep ΔEphb2 mice fed the CDAA-HFD for 12 weeks, stained with antibodies against EphB2 (red) and Pan-CK (green), with DAPI counterstaining of nuclei in blue. EphB2 fluorescence intensity is reduced in Hep ΔEphb2 mice fed the CDAA-HFD ( B ) Representative images of liver sections of Ephb2 fl/fl (Hep WT ) and Hep Δ Ephb2 mice fed the CDAA-HFD for 12 weeks, stained with H&E and Masson-Trichrome to assess tissue damage and collagen deposition. ( C ) Plasma levels of liver damage enzymes ALT and AST in Ephb2 fl/fl (Hep WT ) and Hep Δ Ephb2 mice fed the CDAA-HFD (n=4-7 mice/group). ( D ) Plasma level of cholesterol and triglycerides in Ephb2 fl/fl (Hep WT ) and Hep ΔEphb2 mice fed the CDAA-HFD ( n = 4-7 mice/group). ( E ) Level of hydroxyproline quantified in the livers of Ephb2 fl/fl (Hep WT ) and Hep Δ Ephb2 mice fed the CDAA-HFD (n = 5-7 mice/group). ( F ) Liver to body weight ratio. ( G ) Liver mRNA levels of fibrotic markers Col1a1, Col1a2, and Acta2 in Ephb2 fl/fl (Hep WT ) and Hep ΔEphb2 mice fed the CDAA-HFD ( n = 4-6 mice/group). ( H ) Liver mRNA levels of inflammatory markers Cd11b, Il-1 β, and Il-6 in Ephb2 fl/fl (Hep WT ) and Hep Δ Ephb2 mice fed the CDAA-HFD ( n = 4-6 mice/group). ( I ) Immunophenotyping of various myeloid cells subsets in the livers of Ephb2 fl/fl (Hep WT ) and Hep Δ Ephb2 mice fed the CDAA-HFD (n=4-7 mice/group). Data are presented as mean ± SEM, ns = not significant.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: Immunofluorescence, Staining, Fluorescence, Clinical Proteomics

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: (A) Dot plot showing Ephb2 gene expression across different hepatic cell types in mouse snRNA-seq analyzed from published dataset GSE212837. (B) Representative immunofluorescence images of human liver sections of healthy and MASH patients stained with antibodies against phosphoEphB2 (Red) and αSMA (green), DAPI counterstain for nuclei is shown in blue. (C) Messenger RNA level of EPHB2 , COL1A2 , and ACTA2 in TGF-β1 stimulated primary human HSCs at various time point (n=3). (D) Western blot showing proteins level of EphB2, aSMA, pSmad2/3 Smad2/3 and Vinculin in TGF-β1 stimulated primary human HSCs at various time point. (E) Messenger RNA level of EPHB2 upon inhibition of canonical TGF-β/SMAD with SIS3 Smad3 inhibitor, SD208 TGFβR1 inhibitor, and SB525334 TGFβR2 inhibitor in TGF-β1-treated primary human HSCs (n=5-7). (F) Effects of silencing Smad2 and Smad3 with Mission®-esiRNA on TGF-β-induced EPHB2 mRNA expression in primary human HSCs (n=3). Non-targeting Mission®-esiRNA was used as control. (G) Accessibility peaks for the transcription factor SMAD3 on the promoter of human EPHB2 locus identified from a publicly available ChIP-seq dataset of TGF-β1 stimulated LX2 cells (GSE38103). (H) Enrichment of SMAD3 binding sites on the promoter of human EPHB2 validated by ChIP-qPCR in TGF-β1 stimulated primary human HSCs. All these data presented as mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by ANOVA ns = not significant.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: Gene Expression, Immunofluorescence, Staining, Western Blot, Inhibition, esiRNA, Expressing, Control, ChIP-sequencing, Binding Assay, ChIP-qPCR

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: ( A ) Messenger RNA level of EPHB2 , and ACTA2 in TGF-β2 (10ng/ml) stimulated primary human HSCs at various time point (n=3). ( D ) Western blot showing proteins level of EphB2, α SMA , pSmad2/3 Smad2/3 and Vinculin in TGF-β2 stimulated primary human HSCs at various time point. ( C ) Messenger RNA level of EPHB2 , and ACTA2 in TGF-β3 stimulated primary human HSCs at various time point (n=3 biological replicate). ( D ) Western blot showing proteins level of EPHB2, αSMA, pSMAD2/3, SMAD2/3 and Vinculin in TGF-β3 (10ng/ml) stimulated primary human HSCs at various time point. Data are presented as mean ± SEM, ns = not significant, * p < 0.05, ** p < 0.01, *** p <0.001, **** p <0.0001 by ANOVA.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: Western Blot

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: ( A ) Messenger RNA level of EPHB2 and ( B ) ACTA2 and COL1A1 mRNA levels measured in different HSCs lines treated with Mission® EPHB2 -esiRNA (SIGMA), stimulated with TGF-β1 at 10ng/ml and normalized to the same HSCs lines treated with nontargeting Mission®-esiRNA (n=5). ( C ) Representative immunofluorescence images of TGF-β1-treated HSCs transfected with EPHB2 -siRNA and non-targeted -siRNA, stained with antibodies against EphB2 (green) and αSMA (red). ( D ) Western blot gels images showing the effects of silencing EPHB2 with Mission®-esiRNA on TGF-β1-induced EphB2, αSMA, Smurf2, and Vinculin proteins levels in primary human HSCs for 48 hours. ( E ) Potential protein-protein interaction between EphB2 and TGFBRII detected by immunoprecipitation of EphB2 in TGF-β1 stimulated human HSCs followed by western blot of EphB2 and TGFBRII. ( F ) Proximity ligation assay data of EphB2 and TGFβRII interaction in HSCs. ( G ) Western blot gels images showing the effects of silencing EPHB2 with Mission®-esiRNA on TGF-β1-induced EphB2, pSmad2/3, Smad2/3, pAKT-Ser473, AKT, and Vinculin proteins levels in primary human HSCs for 1 hour. ( H ) TGF-β1-stimulated EPHB2-knocked down and control HSCs were transduced with AAV8 expressing luciferase under the control of the TGFβ-SBE-promoter reporter. TGFβ-induced SMAD activation in HSCs for 3 hours was assessed by measuring luciferase activity (n=3). ( I ) Heatmap depicting the top 40 differentially expressed genes in bulk RNA sequencing of TGF-β1-stimulated HSCs lines treated with nontargeting-esiRNA and EPHB2 -esiRNA. ( J ) Gene ontology analysis of the top significantly downregulated or upregulated pathways in RNA sequencing data in I. ( K ) Live cells imaging and ( L ) quantification of HSCs migration in non-targeted and EPHB2 -esiRNA transfected HSCs lines at various time point. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01 by Student’s t test ns = not significant.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: esiRNA, Immunofluorescence, Transfection, Staining, Western Blot, Immunoprecipitation, Proximity Ligation Assay, Control, Transduction, Expressing, Luciferase, Activation Assay, Activity Assay, RNA Sequencing, Imaging, Migration

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: (A) Messenger RNA levels of SMAD7, SMURF1 , and SMURF2 measured in different HSCs lines treated with Mission® EPHB2 -esiRNA (SIGMA), stimulated with TGF-β1 at 10ng/ml and normalized to the same HSCs lines treated with nontargeting Mission®-esiRNA. (B) Quantitative PCR validation of the top fibrogenic genes upregulated in the RNA sequencing data set of TGF-β1-stimulated control HSCs compared to TGF-β1-stimulated EPHB2 -knocked down HSCs (n=4/group * p < 0.05, ** p < 0.01, *** p <0.001, **** p <0.0001 by ANOVA).

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: esiRNA, Real-time Polymerase Chain Reaction, Biomarker Discovery, RNA Sequencing, Control

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: Paralogous relationships between differentially altered kinases in TGF-β1 stimulated controls and EPHB2 knocked down human HSCs. These HSCs were stimulated with TGF-β1 for 48h. CDKs and MAPKs families of Serine/Threonine kinases were significantly affected by knocking down EPHB2 in TGF-β1 stimulated HSCs.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques:

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: ( A ) Representative images of liver sections of Ephb2 fl/fl (HSC WT ) and HSC ΔEphb2 mice fed the CDAA-HFD for 8 weeks, stained with H&E and Masson-Trichrome. ( B ) Level of hydroxyproline quantified in the livers of Ephb2 fl/fl (HSC WT ) and HSC ΔEphb2 mice fed the CDAA-HFD (n = 5-7 mice/group, ** p < 0.01 by Student’s t test). ( C ) Plasma levels of liver damage enzymes ALT and AST in Ephb2 fl/fl (HSC WT ) and HSC ΔEphb2 mice fed the CDAA-HFD (n=5-7 mice/group). ( D ) Plasma level of cholesterol and triglycerides in Ephb2 fl/fl (HSC WT ) and HSC ΔEphb2 mice fed the CDAA-HFD ( n = 5-7 mice/group). ( E ) Liver mRNA levels of Col1a1, Col3a1, Col6a2, and Acta2 in Ephb2 fl/fl (HSC WT ) and HSC ΔEphb2 mice fed the CDAA-HFD ( n = 5-7 mice/group, * p < 0.05, ** p < 0.01, by Student t test). ( F ) Liver mRNA levels of Cd11b, Il-1 β, and Il-6 in Ephb2 fl/fl (HSC WT ) and HSC ΔEphb2 mice fed the CDAA-HFD ( n = 5-7 mice/group). ( G ) Heatmap depicting the top 40 differentially expressed genes from bulk RNA sequencing of livers tissues of Ephb2 fl/fl (HSC WT ) and HSC ΔEphb2 mice fed the CDAA-HFD. ( H ) Gene ontology analysis of the top significantly downregulated or upregulated pathways in RNA sequencing data in G. ( I ) Percentage of subsets of myeloid cells population in the livers of Ephb2 fl/fl (HSC WT ) and HSC ΔEphb2 mice fed the CDAA-HFD (n = 5-7 mice/group). Data are presented as mean ± SEM, ns = not significant.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: Staining, Clinical Proteomics, RNA Sequencing

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: EphB2 fl/fl mice (Cre-) were crossed with the tamoxifen-inducible Col1a2Cre ERT2 mice to generate Col1a2Cre ERT2 /Ephb2 fl/fl (Cre+) lines. ( A ) Representative images of liver sections of Cre- and Cre+ mice fed the GAN diet for 26 weeks, stained with H&E and Masson-Trichrome. ( B ) Level of hydroxyproline quantified in the livers of Cre- and Cre+ mice fed the GAN diet (n = 5 mice/group, * p < 0.05 by Student’s t test). ( C ) Plasma levels of liver damage enzymes ALT and AST in Cre- and Cre+ mice fed the GAN diet (n=5 mice/group). ( D ) Plasma level of cholesterol and triglycerides in Cre- and Cre+ mice fed the GAN diet for 26 weeks ( n = 5 mice/group, * p < 0.05 by Student’s t test). ( E ) liver to body weight ratio of the livers of Cre- and Cre+ mice fed the GAN diet (n = 5 mice/group). ( F ) Liver mRNA levels of Acta2, Col1a1, Col1a2, and Timp1 in Cre- and Cre+ mice fed the GAN diet ( n = 5 mice/group, * p < 0.05 by Student t test). ( G ) Liver mRNA levels of Cd11b, Il-1 β, IL-6 and Cxcl-10 in Cre- and Cre+ mice fed the GAN diet ( n = 5 mice/group). ( H ) Immunophenotyping of macrophages subset (CD11b, Ly6C, F4/80, MHC-II) dendritic cells (cDC1) and neutrophils (PMN) in the liver of Cre- and Cre+ mice fed the GAN diet (n = 5 mice/group, * p < 0.05 by Student’s t test). Data are presented as mean ± SEM, ns = not significant.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: Staining, Clinical Proteomics

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: EphB2 fl/fl mice (Cre-) were crossed with the tamoxifen-inducible Col1a2Cre ERT2 mice to generate Col1a2Cre ERT2 /Ephb2 fl/fl (Cre+) lines. ( A ) Representative images of liver sections of Cre- and Cre+ mice treated with CCl 4 for 4 weeks, stained with H&E and Masson-Trichrome to assess tissue damage and collagen deposition. ( B ) Level of hydroxyproline quantified in the livers of Cre- and Cre+ mice treated with CCl 4 for 4 weeks (n = 6-9 mice/group, * p < 0.05 by Student’s t test). ( C ) Plasma levels of liver damage enzymes ALT and AST in Cre- and Cre+ mice treated with CCl 4 for 4 weeks (n =6-10 mice/group). ( D ) Liver mRNA levels of Col1a1, Fn, Acta2, and Timp1 in Cre- and Cre+ mice receiving CCl4 for 4 weeks ( n = 6-9 mice/group, * p < 0.05, ** p < 0.01 by Student t test). ( E ) Liver mRNA levels of Cd11b, Ccl-5, Cxcl-10, and Il-1 β in Cre- and Cre+ mice receiving CCl 4 for 4 weeks ( n = 6-9 mice/group, * p < 0.05, ** p < 0.01 by Student t test). ( F ) Immunophenotyping of T cell (CD3), B cells (CD19), and macrophages subset (Ly6C, F4/80, PMN,) in the livers of Cre- and Cre+ mice receiving CCl 4 for 4 weeks (n = 6-9 mice/group, * p < 0.05 by Student’s t test). Data are presented as mean ± SEM, ns = not significant.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: Staining, Clinical Proteomics

Journal: bioRxiv

Article Title: EPHB2 promotes diet-induced MASH liver fibrosis

doi: 10.1101/2025.11.19.688968

Figure Lengend Snippet: ( A ) Experimental design used to delete EphB2 in liver mesenchymal cells when fibrosis is already present in Col1a2Cre ERT2 /Ephb2 WT ( Col WT ) and Col1a2Cre ERT2 /Ephb2 fl/fl ( Col Δ Ephb2 ) fed the CDAA-HFD. ( B ) Liver sections of Col WT and Col ΔEphb2 mice fed the CDAA-HFD for 12 weeks were stained with H&E and Masson-Trichrome to assess tissue damage and collagen deposition. ( C ) Plasma levels of liver damage enzymes ALT and AST in Col WT and Col Δ Ephb2 mice fed the CDAA-HFD (n=4-5 mice/group). ( D ) Plasma level of cholesterol and triglycerides in Col WT and Col Δ Ephb2 mice fed the CDAA-HFD for 12 weeks ( n = 4-5 mice/group). ( E ) Level of hydroxyproline quantified in the livers of Col WT and Col Δ Ephb2 mice fed the CDAA-HFD (n = 4-5 mice/group). ( F ) Liver mRNA level of Ephb2 in Col WT and Col Δ Ephb2 mice fed the CDAA-HFD ( n = 5 mice/group, * p < 0.05 by Student t test). ( G) Liver mRNA levels of Acta2, Col1a1, Col1a2, Col3a1, Col6a2, and Pdgfr β in Col WT and Col Δ Ephb2 mice fed the CDAA-HFD ( n = 5 mice/group, * p < 0.05, ** p < 0.01 by Student t test). ( H ) Liver mRNA levels of Cd11b, IL-6 and Tnf- α in Col WT and Col Δ Ephb2 mice fed the CDAA-HFD ( n = 5 mice/group). Data are presented as mean ± SEM, ns = not significant.

Article Snippet: The cells were then washed three times with TBS-T, blocked for one hour in blocking solution (5% donkey serum; 0.1% triton-X-100 in TBS) and incubated overnight at 4 °C with a primary antibody against EPHB2 (AF467, R&D System), and α-SMA (clone 1A4, R&D system; 1:200 in blocking solution).

Techniques: Staining, Clinical Proteomics

Journal: Biological psychiatry

Article Title: Neuroprotective Peptide NAPVSIPQ Antagonizes Ethanol Inhibition of L1 Adhesion by Promoting the Dissociation of L1 and Ankyrin-G

doi: 10.1016/j.biopsych.2019.08.020

Figure Lengend Snippet: NAP activation of EphB2 phosphorylation of L1. (A) Effect of NAP on association of L1 with EphB2. L1 was immunoprecipitated with mAb 5G3 from 2A2-L1s cells in the absence and presence of 10−9 M NAP, and coimmunoprecipitated proteins were separated and blotted with antibodies to L1 and EphB2. Densities of EphB2 bands were normalized to those for L1, and values for NAP treatment were expressed as a percentage of control values. Shown is the mean ± SEM % increase in L1 association with EphB2 following NAP treatment derived from 8-9 independent experiments; *t=3.17,*p <0.05, n = 8. (B) Dose-dependent stimulation by NAP of tyrosine phosphorylation of FIGQY-peptide by recombinant EphB2 (F = 2.45, p<0.05). pY levels following NAP treatment were normalized to control values (0 NAP). Shown is the mean ± SEM % increase in pY levels following treatment with the indicated concentrations of NAP derived from 6-9 independent experiments (C) Stimulation of EphB2 phosphorylation of L1 by 10−9 M NAP and P7A-NAP (P7A), but not by 10−9 M I6A-NAP (I6A), SAL, or octanol (Oct). Shown is a representative gel and densitometric analysis from 7 independent experiments. pY levels for each drug treatment were normalized to values obtained in the absence of drugs (Control) (F = 2.84, p<0.05); **t = 4.01, **p = 0.0070; *t= 3.01, *p=0.0235, n=7.

Article Snippet: , mAB against EphB2 , Santa Cruz Biotech , SC-130752, RRID: AB_2099957 , .

Techniques: Activation Assay, Phospho-proteomics, Immunoprecipitation, Control, Derivative Assay, Recombinant

Journal: Biological psychiatry

Article Title: Neuroprotective Peptide NAPVSIPQ Antagonizes Ethanol Inhibition of L1 Adhesion by Promoting the Dissociation of L1 and Ankyrin-G

doi: 10.1016/j.biopsych.2019.08.020

Figure Lengend Snippet: Effect of EphB2 knockdown on NAP antagonism of ethanol inhibition of L1 adhesion. (A) 2A2-L1s cells were treated with an EphB2 siRNA or a scrambled siRNA. L1 and actin were used as loading controls, and densities of protein bands were normalized to those obtained in untreated cells (Control). Shown is a representative gel and mean ± SEM % changes in EphB2 expression derived from 9 independent experiments (F=9.33, p< 0.001); * t =1.07, p <0.0.0044, n=10. (B) EphB2 siRNA specifically reduced while scrambled siRNA had no effect on the phosphorylation of FIGQY-peptide from lysates of 2A2-L1s cells treated with 10−12 M NAP. Values for pY in NAP-treated siRNA-treated cells were normalized to values in control cells that were not treated with NAP (F = 2.77, p < 0.05); *t = 2.47, p =0.04, n=8; t = 2.54, p=0.039, n =8. (C) EphB2-siRNA specifically reduced while scrambled siRNA had no effect on NAP antagonism of ethanol inhibition of L1 adhesion. Values for ethanol inhibition of L1 adhesion in the presence of NAP were normalized to values obtained in the absence of NAP (38.6 ± 6.9%) (F = 7.17, p < 0.0001); t= 9.30, ***p =0.0000, n = 9; t = 7.41, ***p = 0.0001, n =9).

Article Snippet: , mAB against EphB2 , Santa Cruz Biotech , SC-130752, RRID: AB_2099957 , .

Techniques: Knockdown, Inhibition, Control, Expressing, Derivative Assay, Phospho-proteomics

Journal: Biological psychiatry

Article Title: Neuroprotective Peptide NAPVSIPQ Antagonizes Ethanol Inhibition of L1 Adhesion by Promoting the Dissociation of L1 and Ankyrin-G

doi: 10.1016/j.biopsych.2019.08.020

Figure Lengend Snippet: NAP antagonizes ethanol inhibition of L1 adhesion by activating EphB2 phosphorylation of L1-Y1229, leading to the dissociation of L1 from ankyrin-G and the spectrin-actin cytoskeleton. L1 is sensitive to ethanol only when it is associated with ankyrin-G.

Article Snippet: , mAB against EphB2 , Santa Cruz Biotech , SC-130752, RRID: AB_2099957 , .

Techniques: Inhibition, Phospho-proteomics

Journal: Biological psychiatry

Article Title: Neuroprotective Peptide NAPVSIPQ Antagonizes Ethanol Inhibition of L1 Adhesion by Promoting the Dissociation of L1 and Ankyrin-G

doi: 10.1016/j.biopsych.2019.08.020

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: , mAB against EphB2 , Santa Cruz Biotech , SC-130752, RRID: AB_2099957 , .

Techniques: Recombinant, Protein Kinase Assay, Kinase Assay, Plasmid Preparation, Sequencing, Software, Transfection, Construct, Expressing