b16f10 cell line  (ATCC)

Journal: Redox Biology

Article Title: Combination of YAP inhibition and photodynamic therapy induces dual DNA damage and activates STING pathway to enhance immunotherapy in uveal melanoma

doi: 10.1016/j.redox.2025.103965

Figure Lengend Snippet: Activation of the cGAS-STING pathway by HANP/VP via YAP-inhibition and PDT in vitro. (A) The phosphorylation levels of representative proteins of the cGAS-STING pathway were analyzed using Western blot ( left ) and quantified ( right ). After HANP/VP treatment without laser irradiation, the pSTING/STING, pTBK1/TBK1, and pIRF3/IRF3 ratio were significantly increased compared to the untreated cells; the phosphorylation levels were further increased after laser irradiation, n = 3/group. (B) qPCR was used to detect IFN-β1 mRNA levels. Compared with the control group, HANP/VP without Laser treatment significantly increased IFN-β1 expression in B16F10 cells, while HANP/VP combined with laser irradiation further elevated IFN-β1 mRNA levels. When cells were pretreated with ethidium bromide (EB, 120 ng/mL), IFN-β1 mRNA levels decreased in the HANP/VP with Laser irradiation group. Following H151 treatment, IFN-β1 mRNA levels decreased to control group levels regardless of laser irradiation status. n = 3/group. Statistical significance was determined by one-way ANOVA (∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: The mouse melanoma cell line B16F10 was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Activation Assay, Inhibition, In Vitro, Phospho-proteomics, Western Blot, Irradiation, Control, Expressing

Journal: The American Journal of Pathology

Article Title: Inhibiting the Secreted RGDKGE Collagen Peptide Selectively Controls CD8 + T-Cell Migration on Denatured Collagen-IV and Enhances Their Accumulation in Tumors

doi: 10.1016/j.ajpath.2025.09.008

Figure Lengend Snippet: CD8 + T cells control tumor growth following targeting the RGDKGE collagen peptide. C57BL/6 mice were treated intraperitoneally with function-blocking anti–CD8α-depleting antibody (BP0061) or non-specific control antibodies. A: Example of flow cytometry analysis for CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). B: Quantification of CD8 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD8 + T cells from four mice per group. C: Example of flow cytometry analysis for CD4 + T cells from spleens of mice treated with non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). D: Quantification of CD4 + T cells from spleens of mice treated with either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8). Data represent CD4 + T cells from four mice per group. E: Examples of CD8 + T cells (red) in B16F10 tumor section from either non-specific control antibody (Ab Cont) or anti–CD8α-depleting antibody (Anti-CD8) treated mice. F: Quantification of the mean CD8 + T-cell count from four different tumors from each treatment group using five to seven ×200 microscopic fields from each tumor. Data represent CD8 + T-cell counts from four different tumors from each treatment group. G: Quantification of B16F10 tumor size from control antibody-depleted (Ab Cont) or CD8-depleted (Anti-CD8) mice over 14 days. Data points represent tumor volume from four mice per condition. H: Quantification of B16F10 tumor size from control antibody-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody [monoclonal antibody (Mab) XL313] over 14 days. Data points represent tumor volume from eight mice per condition. I: Quantification of B16F10 tumor size from CD8-depleted mice treated with non-specific control antibody (Ab Cont) or anti-RGDKGE antibody (Mab XL313) over 14 days. Data points represent tumor volume from seven to eight mice per condition. Data are given as means ± SEM ( B , D , and F – I ). ∗ P < 0.05, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Scale bars = 50 μm ( E ). SSC, side scatter.

Article Snippet: B16F10 murine melanoma cells were obtained from ATCC (Manassas, VA).

Techniques: Control, Blocking Assay, Flow Cytometry, Cell Characterization

Journal: The American Journal of Pathology

Article Title: Inhibiting the Secreted RGDKGE Collagen Peptide Selectively Controls CD8 + T-Cell Migration on Denatured Collagen-IV and Enhances Their Accumulation in Tumors

doi: 10.1016/j.ajpath.2025.09.008

Figure Lengend Snippet: The RGDKGE collagen peptide alters CD8 + T-cell migration on denatured (Den-Coll-IV), but not native (Nat-Coll-IV), collagen-IV. Cell lysates were prepared from CD8 + T cells isolated from B16F10 tumor-bearing mice or from Jurkat T cells. A and B: Western blot analysis for CD8 and β3 integrin protein in T cells isolated from B16F10 tumor-bearing mice ( A ) and Jurkat T cells ( B ). C and D: Quantification of CD8 + T-cell migration on denatured collagen-IV ( C ) or native collagen-IV ( D ) in the presence of control peptide (CP) or RGDKGE collagen peptide 2 (P2). Data represent change in cell migration from three independent experiments with control CP treatment set to 100% for comparison. E and F: Quantification of Jurkat T-cell migration on denatured collagen-IV ( E ) or native collagen-IV ( F ) in the presence of CP or P2. Data represent change in cell migration from three independent experiments with control CP treatment set to 100% for comparison. G and H: Quantification of CD8 + T-cell migration on denatured collagen-IV ( G ) and native collagen-IV ( H ) following incubation with P2 and either control antibody (Ab Cont) or anti-RGDKGE antibody [monoclonal antibody (Mab) XL313]. G: Data represent change in cell migration from five independent experiments with Ab control treatment set to 100% for comparison. H: Data represent change in cell migration from four independent experiments with Ab control treatment set to 100% for comparison. I and J: Quantification of Jurkat T-cell migration on denatured collagen-IV ( I ) and native collagen-IV ( J ) following incubation with P2 and Ab Cont or anti-RGDKGE antibody (Mab XL313). I: Data represent change in cell migration from five independent experiments with Ab control treatment set to 100% for comparison. J: Data represent change in cell migration from three independent experiments with Ab control treatment set to 100% for comparison. K: Western blot analysis of the secreted RGDKGE collagen peptide in 10× concentrated serum-free control medium (Cont Media) or B16F10 conditioned medium (B16F10 CM). L and M: Quantification of CD8 + T-cell migration on denatured collagen-IV ( L ) and Jurkat T cells ( M ) following incubation with serum-free B16F10 cell CM and Ab Cont or anti-RGDKGE antibody (Mab XL313). Data represent change in cell migration from three independent experiments with Ab control treatment set to 100% for comparison. Data are given as means ± SEM ( C – J , L , and M ). ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: B16F10 murine melanoma cells were obtained from ATCC (Manassas, VA).

Techniques: Migration, Isolation, Western Blot, Control, Comparison, Incubation

Journal: The American Journal of Pathology

Article Title: Inhibiting the Secreted RGDKGE Collagen Peptide Selectively Controls CD8 + T-Cell Migration on Denatured Collagen-IV and Enhances Their Accumulation in Tumors

doi: 10.1016/j.ajpath.2025.09.008

Figure Lengend Snippet: The RGDKGE collagen peptide (P2) alters F-actin polarization and CD8 + T-cell infiltration of B16F10 tumors in vivo . A: Example of Jurkat T-cell F-actin polarization ( arrows ) in control peptide (CP) and P2 treated cells seeded onto denatured collagen-IV (Den-Coll-IV). B: Quantification of the mean percentage of F-actin polarized cells per ×400 field in control (CP) and P2 treated Jurkat T cells seeded onto denatured collagen-IV. Data represent percentage of F-actin polarized T cells from three independent experiments calculated from 10 ×400 fields per condition. C: Quantification of the mean percentage of F-actin polarized cells per ×400 field in control antibody and monoclonal antibody (Mab) XL313 treated Jurkat T cells seeded onto denatured collagen-IV in the presence of P2. Data represent percentage F-actin polarized T cells from three independent experiments calculated from 10 ×400 fields per condition. D: Example of F-actin polarization ( arrows ) in CP and P2 treated CD8 + T cells isolated from B16F10 tumor-bearing mice seeded onto denatured collagen-IV. E: Quantification of the mean percentage of F-actin polarized cells per ×400 field in control (CP) and P2 treated CD8 + T cells seeded onto denatured collagen-IV. Data represent percentage of F-actin polarized T cells from two independent experiments calculated from 10 ×400 fields per condition. F: Quantification of the mean percentage of F-actin polarized cells per ×400 field in control antibody and Mab XL313 treated CD8 + T cells seeded onto denatured collagen-IV in the presence of P2. Data represent mean percentage of F-actin polarized CD8 + T cells from four independent experiments calculated from 10 ×400 fields per condition. G: Example of B16F10 melanoma costained with anti-denatured collagen-specific antibody D93 (green) and endothelial cell marker anti-CD31 (red). H: Quantification of the mean CD8 + T cells within single-cell suspensions of B16F10 melanomas growing in C57BL/6 mice. Data represent percentage of CD8 + T cells from eight mice per condition. I: Working model of how the secreted RGDKGE collagen peptide may control CD8 + T-cell migration through denatured collagen-IV–rich microenvironments. Data are given as means ± SEM ( B , C , E , F , and H ). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Scale bars = 20 μm ( A , D , and G ).

Article Snippet: B16F10 murine melanoma cells were obtained from ATCC (Manassas, VA).

Techniques: In Vivo, Control, Isolation, Marker, Single Cell, Migration