Journal: American Journal of Translational Research

Article Title: Overexpression of B7-H3 as an opportunity for targeted therapy in head and neck cancers

doi:

Figure Lengend Snippet: Characterization of Fc-tagged B7-H3 scFvs and the construction of anti-B7-H3×CD3 biAbs. A. Validation of the binding of Fc-tagged B7-H3 scFvs to FADU cells was conducted by immunofluorescence, and the arrows indicated Fc-tagged B7-H3 scFvs binding to FADU cells. B. Schematic diagram of the anti-B7-H3×CD3 biAb expression vector. C. SDS-PAGE analysis of purified anti-B7-H3×CD3 biAbs. The molecular weight was approximately 65 kD. Scale bar: 50 μm.

Article Snippet: The slides were incubated with the primary antibody or Fc-tagged B7-H3 scFv for 1 hour, followed by incubation with the secondary antibody for 1 h. Finally, the slides were stained sequentially with a Cy3-conjugated secondary antibody (Proteintech) and 4’,6-diamidino-2-phenylindole (DAPI) (Beyotime).

Techniques: Binding Assay, Immunofluorescence, Expressing, Plasmid Preparation, SDS Page, Purification, Molecular Weight

Journal: American Journal of Translational Research

Article Title: Overexpression of B7-H3 as an opportunity for targeted therapy in head and neck cancers

doi:

Figure Lengend Snippet: B7-H1, B7-H3, B7-H4 and B7-H5 expression in various head and neck tissues. A. Box plots of B7-H1, B7-H3, B7-H4 and B7-H5 mRNA expression in head and neck tissues from TCGA (*P<0.05). B. IHC analysis of B7-H1, B7-H3, B7-H4 and B7-H5 expression was evaluated in normal head and neck tissues and HNC tissues.

Article Snippet: The slides were incubated with the primary antibody or Fc-tagged B7-H3 scFv for 1 hour, followed by incubation with the secondary antibody for 1 h. Finally, the slides were stained sequentially with a Cy3-conjugated secondary antibody (Proteintech) and 4’,6-diamidino-2-phenylindole (DAPI) (Beyotime).

Techniques: Expressing

Journal: American Journal of Translational Research

Article Title: Overexpression of B7-H3 as an opportunity for targeted therapy in head and neck cancers

doi:

Figure Lengend Snippet: Analysis of B7-H3 expression in various head and neck tissues. A. IHC analysis of B7-H3 expression in four different histological subtypes. B and D. IHC analysis was conducted in HNC tissues with different intensities of B7-H3 and densities of CD3 staining. The red arrows indicated B7-H3 positive staining, and the green triangles indicated CD3 positive staining. C. Kaplan-Meier curve for the HNC-specific survival of HNC patients with low or high B7-H3 expression. The duration of survival was measured from the first postoperative day to the time of death or at 107 months. Scale bar: 100 μm.

Article Snippet: The slides were incubated with the primary antibody or Fc-tagged B7-H3 scFv for 1 hour, followed by incubation with the secondary antibody for 1 h. Finally, the slides were stained sequentially with a Cy3-conjugated secondary antibody (Proteintech) and 4’,6-diamidino-2-phenylindole (DAPI) (Beyotime).

Techniques: Expressing, Staining

Journal: American Journal of Translational Research

Article Title: Overexpression of B7-H3 as an opportunity for targeted therapy in head and neck cancers

doi:

Figure Lengend Snippet: Association of B7-H3 expression levels in HNC samples with clinicopathological parameters

Article Snippet: The slides were incubated with the primary antibody or Fc-tagged B7-H3 scFv for 1 hour, followed by incubation with the secondary antibody for 1 h. Finally, the slides were stained sequentially with a Cy3-conjugated secondary antibody (Proteintech) and 4’,6-diamidino-2-phenylindole (DAPI) (Beyotime).

Techniques: Expressing

Journal: American Journal of Translational Research

Article Title: Overexpression of B7-H3 as an opportunity for targeted therapy in head and neck cancers

doi:

Figure Lengend Snippet: Logistic regression analysis of high B7-H3 expression in HNCs

Article Snippet: The slides were incubated with the primary antibody or Fc-tagged B7-H3 scFv for 1 hour, followed by incubation with the secondary antibody for 1 h. Finally, the slides were stained sequentially with a Cy3-conjugated secondary antibody (Proteintech) and 4’,6-diamidino-2-phenylindole (DAPI) (Beyotime).

Techniques: Expressing

Journal: American Journal of Translational Research

Article Title: Overexpression of B7-H3 as an opportunity for targeted therapy in head and neck cancers

doi:

Figure Lengend Snippet: B7-H3 expression in different HNC cell lines. A and B. Immunofluorescence and the corresponding semiquantitative image analysis showed different levels of B7-H3 expression in HNC cell lines. The red fluorescence was stained with Cy3-conjugated secondary antibody (Proteintech, 1:500), and the nucleus was stained with DAPI (Beyotime, 1:10). C (original western blotting image in the Figure S3A) and D. Western blotting and the corresponding semiquantitative image analysis showed different levels of B7-H3 expression in HNC cell lines. Data was represented as the mean ± SD. (-, negative; +, weak; ++, mild; +++, strong). (*P<0.05, **P<0.01, ***P<0.001). Scale bar: 20 μm.

Article Snippet: The slides were incubated with the primary antibody or Fc-tagged B7-H3 scFv for 1 hour, followed by incubation with the secondary antibody for 1 h. Finally, the slides were stained sequentially with a Cy3-conjugated secondary antibody (Proteintech) and 4’,6-diamidino-2-phenylindole (DAPI) (Beyotime).

Techniques: Expressing, Immunofluorescence, Fluorescence, Staining, Western Blot

Journal: American Journal of Translational Research

Article Title: Overexpression of B7-H3 as an opportunity for targeted therapy in head and neck cancers

doi:

Figure Lengend Snippet: Cytotoxicity analysis of HNCs treated with anti-B7-H3×CD3 biAbs in vitro. A. Specific lysis of FADU cells treated with anti-B7-H3×CD3 biAbs for different durations. Obvious cytolysis was observed in the anti-B7-H3×CD3 biAb groups after 24 hours, while no cytolysis was observed in the PBS groups. B. Compared to the PBS + T-cells groups, anti-B7-H3×CD3 biAbs + T-cells groups showed statistically significant viability after a 48-hour incubation period. C. Specific lysis of FADU cells treated with different E:T ratios and anti-B7-H3×CD3 biAb concentrations. D. Cell growth inhibition curves for different HNC cell lines with different levels of B7-H3 expression. The IC50 values were shown on each curve. Dotted boxes indicated the distribution and growth status of FADU cells in the wells. The red arrows indicated T-cells adhering slightly to the surface of FADU cells, and the green triangles indicated the lysis of FADU cells. Data was represented as the mean ± SD. (n.s. not significant, *P<0.05, **P<0.01, ***P<0.001). Relative absorbance value: RAU. Scale bar: 50 μm.

Article Snippet: The slides were incubated with the primary antibody or Fc-tagged B7-H3 scFv for 1 hour, followed by incubation with the secondary antibody for 1 h. Finally, the slides were stained sequentially with a Cy3-conjugated secondary antibody (Proteintech) and 4’,6-diamidino-2-phenylindole (DAPI) (Beyotime).

Techniques: In Vitro, Lysis, Incubation, Inhibition, Expressing

Journal: American Journal of Translational Research

Article Title: Overexpression of B7-H3 as an opportunity for targeted therapy in head and neck cancers

doi:

Figure Lengend Snippet: Cytotoxicity analysis of HNC xenografts in mice treated with anti-B7-H3×CD3 biAbs in vivo. A. In vivo bioluminescence imaging of all 10 mice with luciferase-expressing FADU tumors on day 7 (before treatment), day 25 and day 34 (after treatment) was shown. B. Tumor growth curves for FADU tumors. C. Survival curves for mice engrafted with PBS-treated tumors and anti-B7-H3×CD3 biAb-treated tumors. D. Representative images of B7-H3 and CD3 IHC staining in tumor sections. The red arrows indicated positive CD3 staining, the green triangles indicated the tumor cell compartment, and the black stars indicated necrotic and disintegrating tumor cells. E. Western blotting showing expression of B7-H3 and tubulin in tumor sections (original western blotting image is in the Figure S3B). Scale bar: 50 μm.

Article Snippet: The slides were incubated with the primary antibody or Fc-tagged B7-H3 scFv for 1 hour, followed by incubation with the secondary antibody for 1 h. Finally, the slides were stained sequentially with a Cy3-conjugated secondary antibody (Proteintech) and 4’,6-diamidino-2-phenylindole (DAPI) (Beyotime).

Techniques: In Vivo, Imaging, Luciferase, Expressing, Immunohistochemistry, Staining, Western Blot