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

Journal: Glycobiology

Article Title: Editor's Choice Functional inactivation of oligosaccharyltransferase a isoform suppresses tumor metastasis

doi: 10.1093/glycob/cwag003

Figure Lengend Snippet: Syngeneic model. (A) Schematic representation of the experimental model. When cancer cell-derived EVs that contain Cre25nt mRNA were incorporated into recipient cells, the mRNA may be translated to Cre recombinase, leading to expression of tdTomato protein. (B and C) RT-PCR and western blot analysis of Cre25nt mRNA (B) and Cre protein (C) in wild-type (WT) B16-F10 cells, Cre25nt cells, and small EVs secreted from these cells. (D) Expression of tdTomato (red) in tumors formed by WT cells and Cre25nt cells. tdTomato protein was also detected by anti-red fluorescent protein (RFP) antibody (green). Sections were counterstained with Hoechst 33342 (cyan). Scale bars, 100 μm.

Article Snippet: The mouse melanoma cell line B16-F10 was purchased from American Type Culture Collection (ATCC).

Techniques: Derivative Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Glycobiology

Article Title: Editor's Choice Functional inactivation of oligosaccharyltransferase a isoform suppresses tumor metastasis

doi: 10.1093/glycob/cwag003

Figure Lengend Snippet: Gene-edited knockout of the Stt3a and Stt3b genes in mouse melanoma cells. (A and B) Western blot analysis of wild-type (WT) B16-F10, Stt3a -RE, Stt3a -KO, Stt3b -RE, and Stt3b -KO cells. Samples were treated with or without PNGase F (B). (C) Quantitative real-time PCR analysis of Cre25nt mRNA in small EVs secreted from Stt3a -RE, Stt3a -KO, Stt3b -RE, and Stt3b -KO cells. Average expression of Stt3a -KO, Stt3b -KO were set to 1.0. Data represent the mean ± standard deviation; n = 3.

Article Snippet: The mouse melanoma cell line B16-F10 was purchased from American Type Culture Collection (ATCC).

Techniques: Knock-Out, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Standard Deviation