Journal: Oncotarget

Article Title: Assessment of combined expression of B7-H3 and B7-H4 as prognostic marker in esophageal cancer patients

doi: 10.18632/oncotarget.12628

Figure Lengend Snippet: The panel A. represented the high expression of both B7-H3 and B7-H4 proteins, while panel B. showed, B7-H3 high and B7-H4 low expression, and panel C. revealed B7-H3 low and B7-H4 high expression. The panel D. represented the low expression of both B7-H3 and B7-H4 proteins. The panel E. represented the weak expression of B7-H3 as well as B7-H4 in adjacent normal esophageal tissues. (Scale bar = 100 μm).

Article Snippet: Rabbit anti-human B7-H4 polyclonal antibody (NBP2-30536) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Expressing

Journal: Oncotarget

Article Title: Assessment of combined expression of B7-H3 and B7-H4 as prognostic marker in esophageal cancer patients

doi: 10.18632/oncotarget.12628

Figure Lengend Snippet: Correlation between the clinical parameters and B7-H3 expression in combination with B7-H4 expression in esophageal cancer tissues

Article Snippet: Rabbit anti-human B7-H4 polyclonal antibody (NBP2-30536) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Expressing

Journal: Oncotarget

Article Title: Assessment of combined expression of B7-H3 and B7-H4 as prognostic marker in esophageal cancer patients

doi: 10.18632/oncotarget.12628

Figure Lengend Snippet: Panel A. showed that comparison of the overall survival rate of the patients with B7-H3 high and B7-H4 low vs. patients with both B7-H3 and B7-H4 high expression. Panel B. showed the comparison of the overall survival rate of the patients with both B7-H3 and B7-H4 high expression vs. all the other patients.

Article Snippet: Rabbit anti-human B7-H4 polyclonal antibody (NBP2-30536) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Comparison, Expressing

Journal: Oncotarget

Article Title: Assessment of combined expression of B7-H3 and B7-H4 as prognostic marker in esophageal cancer patients

doi: 10.18632/oncotarget.12628

Figure Lengend Snippet: Prognostic factors analysis based on Cox's proportional hazards model

Article Snippet: Rabbit anti-human B7-H4 polyclonal antibody (NBP2-30536) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques:

Journal: Cell death discovery

Article Title: Counteracting gemcitabine+nab-paclitaxel induced dysbiosis in KRAS wild type and KRAS G12D mutated pancreatic cancer in vivo model.

doi: 10.1038/s41420-023-01397-y

Figure Lengend Snippet: Fig. 2 Immunophenotypical characteristics of mice pancreatic cancer under chemotherapy and/or probiotic treatment. Representative immunohistochemical analyses of Ki67 (×20 magnification), Phospho- γH2A.X (×20 magnification), Arginase 1 (×20 magnification), and AIF (×10 magnification) in BxPC-3 (A) and PANC-1 (B) tumor sections. Scale bar 50 µm.

Article Snippet: Tissue sections were blocked with FBS serum in PBS for 60min and incubated overnight with the following primary antibodies: Ki67 (Thermoscientific, Italy, MA5-14520), Phospho-γH2A.X (Abcam, Italy, ab1174), Arginase-1 (GeneTex, USA, California, GTX109242), Apoptosis-Inducing Factor or AIF (Cell Signaling, USA, Massachusetts, 5318), Myeloperoxidase (Abcam, ab208670), CD41 (Abcam, ab181582), PAX5 (Abcam, ab109443), TER119 (Abcam, ab91113), and Endomucin (Abcam, ab106100).

Techniques: Immunohistochemical staining

Journal: Cell death discovery

Article Title: Counteracting gemcitabine+nab-paclitaxel induced dysbiosis in KRAS wild type and KRAS G12D mutated pancreatic cancer in vivo model.

doi: 10.1038/s41420-023-01397-y

Figure Lengend Snippet: Fig. 4 Role of chemotherapy and/or probiotic treatment on murine small intestine architecture. Representative pictures of hematoxylin/ eosin, Alcian Blue/PAS and Ki67 in BxPC-3 (A) and PANC-1 (B) small intestine sections (×20 magnification). Scale bar 50 µm.

Article Snippet: Tissue sections were blocked with FBS serum in PBS for 60min and incubated overnight with the following primary antibodies: Ki67 (Thermoscientific, Italy, MA5-14520), Phospho-γH2A.X (Abcam, Italy, ab1174), Arginase-1 (GeneTex, USA, California, GTX109242), Apoptosis-Inducing Factor or AIF (Cell Signaling, USA, Massachusetts, 5318), Myeloperoxidase (Abcam, ab208670), CD41 (Abcam, ab181582), PAX5 (Abcam, ab109443), TER119 (Abcam, ab91113), and Endomucin (Abcam, ab106100).

Techniques:

Journal: Scientific reports

Article Title: Interferon-γ in the tumor microenvironment promotes the expression of B7H4 in colorectal cancer cells, thereby inhibiting cytotoxic T cells.

doi: 10.1038/s41598-024-56681-3

Figure Lengend Snippet: Figure 2. IFN-γ stimulation induces immunosuppression in CRC. (A) RNA-seq data of CT26.WT subcutaneous tumors showing the top ten pathways activated in the IFN-γ group (n = 2) compared with the control group (n = 3). The graph shows the category scores as -log10 (p values) determined by Fisher’s exact test. (B) RNA-seq data of CT26.WT subcutaneous tumors showing the top ten upregulated genes in the IFN-γ group (n = 2) compared with the control group (n = 2). The graph shows the category scores as -log10 (p values) determined by Fisher’s exact test. (C) The expression of interferon signaling pathway proteins in CRC cell lines was measured by Western blotting before and after IFN-γ stimulation. (The original images of Western blotting can be found in supplementary Figs. 3–9). (D) Comparison of relative B7h4 RNA expression before and after IFN-γ stimulation in CRC cells, as determined by real-time qPCR. Multiple t tests. (E) Treg cells in the peripheral blood of CT26.WT subcutaneous tumor-bearing IM-c-mice were analyzed by FC. (F) B7H4 protein expression in CT26.WT subcutaneous tumor cells was analyzed by FC. (G) The proportion of CD8+ T cells among CT26.WT subcutaneous tumor cells was determined by FC. (H) B7H4 and CD8 protein expression in CT26.WT subcutaneous tumors was detected by IHC staining. Scale bars, 200 μm. In (E) to (H), *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 by Student’s t test.

Article Snippet: The following primary antibodies were used: anti-VTCN1 (B7H4) (Proteintech, catalog no. 12080-1-AP), anti-PD-L1/CD274 (Proteintech, catalog no. 66248-1-Ig), anti-IRF1 (Proteintech, catalog no. 11335-1-AP), anti-IFNGR1 (Proteintech, catalog no. 10808-1-AP), anti-IFNGR2 (Proteintech, catalog no. 10266- 1-AP), anti-JAK2 (Proteintech, catalog no. 17670-1-AP), anti-STAT1 (Proteintech, catalog no. 10144-2-AP), anti-CD86 (Proteintech, catalog no. 26903-1-AP) and anti-GAPDH (Proteintech, catalog no. 60004-1-Ig).

Techniques: RNA Sequencing, Control, Expressing, Western Blot, Comparison, RNA Expression, Immunohistochemistry

Journal: Scientific reports

Article Title: Interferon-γ in the tumor microenvironment promotes the expression of B7H4 in colorectal cancer cells, thereby inhibiting cytotoxic T cells.

doi: 10.1038/s41598-024-56681-3

Figure Lengend Snippet: Figure 5. IRF1 regulates B7H4 expression. (A) The expression of IRF1 in CRC cells before and after IFN-γ stimulation was determined by Western blotting (the original images of Western blotting can be found in supplementary Figs. 10–14). (B) Representative qPCR results showing the expression of IRF1 before and after IFN-γ stimulation. (C) Relative enrichment of IRF1 binding in the B7H4 promoter, as determined by ChIP‒ qPCR in SW480 and (D) DLD-1 cells. (E) Dual luciferase activity of the B7H4 promoter in SW480 and (F) DLD-1 cells. (G) Changes in B7H4 expression after overexpression of IRF1 (OE-IRF1) or silencing of IRF1 (sh-IRF1) were evaluated by Western blotting in SW480 and (H) DLD-1 cells. (The original images of Western blotting can be found in supplementary Figs. 15, 16). In (B), the data are presented as the means ± SDs. Student’s t test (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001). In (C) to (F), one-way ANOVA (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001).

Article Snippet: The following primary antibodies were used: anti-VTCN1 (B7H4) (Proteintech, catalog no. 12080-1-AP), anti-PD-L1/CD274 (Proteintech, catalog no. 66248-1-Ig), anti-IRF1 (Proteintech, catalog no. 11335-1-AP), anti-IFNGR1 (Proteintech, catalog no. 10808-1-AP), anti-IFNGR2 (Proteintech, catalog no. 10266- 1-AP), anti-JAK2 (Proteintech, catalog no. 17670-1-AP), anti-STAT1 (Proteintech, catalog no. 10144-2-AP), anti-CD86 (Proteintech, catalog no. 26903-1-AP) and anti-GAPDH (Proteintech, catalog no. 60004-1-Ig).

Techniques: Expressing, Western Blot, Binding Assay, Luciferase, Activity Assay, Over Expression

Journal: Scientific reports

Article Title: Interferon-γ in the tumor microenvironment promotes the expression of B7H4 in colorectal cancer cells, thereby inhibiting cytotoxic T cells.

doi: 10.1038/s41598-024-56681-3

Figure Lengend Snippet: Figure 6. The expression and prognostic significance of B7H4 and CD8 in CRC tissues. (A) The expression of B7H4 in CRC tissues was evaluated by IHC staining. Scale bars, 50 μm. (B) The relationship between B7H4 expression and prognosis was confirmed by Kaplan–Meier survival analysis. (C) The expression of CD8 in CRC tissues was evaluated by IHC staining. Scale bars, 50 μm. (D) The relationship between CD8 expression and prognosis was confirmed by Kaplan–Meier survival analysis. Kaplan–Meier survival analysis (**P < 0.01; ****P < 0.0001).

Article Snippet: The following primary antibodies were used: anti-VTCN1 (B7H4) (Proteintech, catalog no. 12080-1-AP), anti-PD-L1/CD274 (Proteintech, catalog no. 66248-1-Ig), anti-IRF1 (Proteintech, catalog no. 11335-1-AP), anti-IFNGR1 (Proteintech, catalog no. 10808-1-AP), anti-IFNGR2 (Proteintech, catalog no. 10266- 1-AP), anti-JAK2 (Proteintech, catalog no. 17670-1-AP), anti-STAT1 (Proteintech, catalog no. 10144-2-AP), anti-CD86 (Proteintech, catalog no. 26903-1-AP) and anti-GAPDH (Proteintech, catalog no. 60004-1-Ig).

Techniques: Expressing, Immunohistochemistry

Journal: Oncology Letters

Article Title: Increased B7-H4 expression during esophageal squamous cell carcinogenesis is associated with IL-6/STAT3 signaling pathway activation in mice

doi: 10.3892/ol.2017.5688

Figure Lengend Snippet: B7-H4 expression, and CD4 + T, CD8 + T and CD11b + macrophage cell infiltration in esophagi of mice, as estimated by immunohistochemistry staining. B7-H4 was expressed in the membrane and cytoplasm of epithelial cells in the mucosa layer. Infiltration of CD4 + T, CD8 + T and CD11b + macrophage cells was predominantly located in the submucosa of esophageal tissues. Magnification, ×400. The arrows indicate the cells that are positive. B7-H4, B7-homolog 4; CD, cluster of differentiation.

Article Snippet: Sections were incubated with primary antibodies against B7-H4 (1:500; GTX62090; GeneTex, Inc., Irvine, CA, USA), CD4 (1:50; orb4241; Biorbyt, Ltd., Cambridge, UK), CD8 (1:50; orb10325; Biorbyt, Ltd.) or CD11b (1:100; cb1211; Abcam, Shanghai, China) at 4°C overnight.

Techniques: Expressing, Immunohistochemistry, Staining, Membrane

Journal: Oncology Letters

Article Title: Increased B7-H4 expression during esophageal squamous cell carcinogenesis is associated with IL-6/STAT3 signaling pathway activation in mice

doi: 10.3892/ol.2017.5688

Figure Lengend Snippet: Association between B7-H4 expression and pathological stage or lymphocytes infiltration in the epithelia of esophageal tissues during the carcinogenesis process.

Article Snippet: Sections were incubated with primary antibodies against B7-H4 (1:500; GTX62090; GeneTex, Inc., Irvine, CA, USA), CD4 (1:50; orb4241; Biorbyt, Ltd., Cambridge, UK), CD8 (1:50; orb10325; Biorbyt, Ltd.) or CD11b (1:100; cb1211; Abcam, Shanghai, China) at 4°C overnight.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Increased B7-H4 expression during esophageal squamous cell carcinogenesis is associated with IL-6/STAT3 signaling pathway activation in mice

doi: 10.3892/ol.2017.5688

Figure Lengend Snippet: Gene expression of B7-H4 and associated molecules in control mice. The expression of genes in the esophagi of control mice did not change obviously from weeks 16 to 28 (B7-H4: P=0.0650; IL-6: P=0.2842; IL-10: P=0.2816; TGF-β, P=0.0743; IFN-γ: P=0.3399; STAT3: P=0.3225; Kruskal-Wallis analysis). mRNA, messenger RNA; Cq, quantification cycle; B7-H4, B7-homolog 4; CD, cluster of differentiation; IL, interleukin; TGF, transforming growth factor; IFN, interferon; STAT3, signal transducer and activator of transcription 3.

Article Snippet: Sections were incubated with primary antibodies against B7-H4 (1:500; GTX62090; GeneTex, Inc., Irvine, CA, USA), CD4 (1:50; orb4241; Biorbyt, Ltd., Cambridge, UK), CD8 (1:50; orb10325; Biorbyt, Ltd.) or CD11b (1:100; cb1211; Abcam, Shanghai, China) at 4°C overnight.

Techniques: Gene Expression, Control, Expressing

Journal: Oncology Letters

Article Title: Increased B7-H4 expression during esophageal squamous cell carcinogenesis is associated with IL-6/STAT3 signaling pathway activation in mice

doi: 10.3892/ol.2017.5688

Figure Lengend Snippet: Gene expression of B7-H4, IL-6, IL-10, TGF-β and STAT3 in the esophagi of 4NQO model mice increased during the carcinogenesis process (*P<0.05, **P<0.01, vs. the control group; Kruskal-Wallis analysis). mRNA, messenger RNA; Cq, quantification cycle; B7-H4, B7-homolog 4; CD, cluster of differentiation; IL, interleukin; TGF, transforming growth factor; IFN, interferon; STAT3, signal transducer and activator of transcription 3; 4NQO, 4-nitroquinoline-1-oxide.

Article Snippet: Sections were incubated with primary antibodies against B7-H4 (1:500; GTX62090; GeneTex, Inc., Irvine, CA, USA), CD4 (1:50; orb4241; Biorbyt, Ltd., Cambridge, UK), CD8 (1:50; orb10325; Biorbyt, Ltd.) or CD11b (1:100; cb1211; Abcam, Shanghai, China) at 4°C overnight.

Techniques: Gene Expression, Control

Journal: Oncology Letters

Article Title: Increased B7-H4 expression during esophageal squamous cell carcinogenesis is associated with IL-6/STAT3 signaling pathway activation in mice

doi: 10.3892/ol.2017.5688

Figure Lengend Snippet: Expression of B7-H4 and associated proteins in control mice. Protein expression in the esophagi of control mice did not change obviously from weeks 16 to 28 (all P>0.05, Kruskal-Wallis analysis). B7-H4, B7-homolog 4; IL, interleukin; STAT3, signal transducer and activator of transcription 3; p-, phosphorylated.

Article Snippet: Sections were incubated with primary antibodies against B7-H4 (1:500; GTX62090; GeneTex, Inc., Irvine, CA, USA), CD4 (1:50; orb4241; Biorbyt, Ltd., Cambridge, UK), CD8 (1:50; orb10325; Biorbyt, Ltd.) or CD11b (1:100; cb1211; Abcam, Shanghai, China) at 4°C overnight.

Techniques: Expressing, Control

Journal: Oncology Letters

Article Title: Increased B7-H4 expression during esophageal squamous cell carcinogenesis is associated with IL-6/STAT3 signaling pathway activation in mice

doi: 10.3892/ol.2017.5688

Figure Lengend Snippet: B7-H4, p-STAT3 and IL-6 protein expression in the esophagi of 4NQO model mice increased during the carcinogenesis process (*P<0.05, **P<0.01 vs. the control group; Kruskal-Wallis analysis). B7-H4, B7-homolog 4; IL, interleukin; STAT3, signal transducer and activator of transcription 3; p-, phosphorylated; 4NQO, 4-nitroquinoline-1-oxide.

Article Snippet: Sections were incubated with primary antibodies against B7-H4 (1:500; GTX62090; GeneTex, Inc., Irvine, CA, USA), CD4 (1:50; orb4241; Biorbyt, Ltd., Cambridge, UK), CD8 (1:50; orb10325; Biorbyt, Ltd.) or CD11b (1:100; cb1211; Abcam, Shanghai, China) at 4°C overnight.

Techniques: Expressing, Control