b16 f10 mouse melanoma tumor 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: Frontiers in Cell and Developmental Biology

Article Title: ERM proteins support perinuclear actin rim formation

doi: 10.3389/fcell.2025.1579946

Figure Lengend Snippet: Perinuclear localization of ERM proteins. Confluent B16–F10 cells induced to migrate in the wound healing assay for 3 h, immunostained for Radixin (A) , Moesin (B) , and Ezrin (C) , along with Lamin B. DNA is stained with Hoechst. The edge of the scratch is in the top region of each micrograph. The nuclei in the orange rectangles are magnified. Scale bar: 20 µm. For quantification, 20–45 cells from each condition were measured in each experiment for the ERM protein signal at the nuclear periphery. The mean intensity was calculated and normalized to control cells. The average mean intensity in three independent experiments ±s.e. is presented. Statistical significance was evaluated by the Student’s t-test, * P < 0.05.

Article Snippet: Mouse melanoma B16-F10 cell line purchased from ATCC was grown as described previously ( ).

Techniques: Wound Healing Assay, Staining, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: ERM proteins support perinuclear actin rim formation

doi: 10.3389/fcell.2025.1579946

Figure Lengend Snippet: Overexpression of ERM proteins increases the intensity of the perinuclear actin rim. (A) Perinuclear actin rim upon overexpression of ERM proteins. Confluent B16-F10 cells over-expressing GFP-fused ERM proteins were induced to migrate in the wound healing assay for 3 h, stained for filamentous actin (Phalloidin), nuclear envelope (Lamin B), and DNA (Hoechst). The edge of the scratch is in the top region of each micrograph. The nuclei in the orange rectangles are magnified on the left side. Scale bar: 20 µm. (B) Quantification of the actin perinuclear rim in ERM overexpressing cells vs. control cells. For quantification, 20–30 cells from each condition were measured for the Phalloidin signal at the nuclear periphery in each experiment. The mean intensity was calculated and normalized to control cells. The average mean intensity in three independent experiments ±s.e. is presented. Statistical significance was evaluated by the Student’s t-test, * P < 0.05, ** P < 0.01.

Article Snippet: Mouse melanoma B16-F10 cell line purchased from ATCC was grown as described previously ( ).

Techniques: Over Expression, Expressing, Wound Healing Assay, Staining, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: ERM proteins support perinuclear actin rim formation

doi: 10.3389/fcell.2025.1579946

Figure Lengend Snippet: ERM proteins support the formation of a perinuclear actin rim. (A) Actin perinuclear rim after KD of ERM proteins. Sub-confluent B16–F10 cells transfected with either control, Radixin, Moesin, or Ezrin siRNA stained for filamentous actin (Phalloidin), nuclear envelope (Lamin B), and DNA (Hoechst). The nuclei in the orange rectangles are magnified on the left side. Scale bar: 20 µm. (B) Quantification of the actin perinuclear rim in siRNA ERM proteins vs. siRNA Control transfected B16–F10 cells. For quantification, in each experiment, 20–30 cells of each transfection were measured for the Phalloidin signal at the nuclear periphery. The mean intensity was calculated and normalized to control cells. The average mean intensity in three independent experiments ±s.e. is presented. Statistical significance was evaluated by the Student’s t-test, ** P < 0.01.

Article Snippet: Mouse melanoma B16-F10 cell line purchased from ATCC was grown as described previously ( ).

Techniques: Transfection, Control, Staining

Journal: International Journal of Molecular Sciences

Article Title: Disulfiram/Copper Combined with Irradiation Induces Immunogenic Cell Death in Melanoma

doi: 10.3390/ijms27020980

Figure Lengend Snippet: Cell viability of MV3 and B16F10 cells treated with DSF/Cu. After MV3 and B16F10 cells were treated with DSF: 0–0.4 µM, Cu: 1 µM for 24 h. Cell viability was assessed using the MTT assay for MV3 ( A ) and B16F10 cells ( B ). Data were generated in triplicate and are presented as mean ± SD. The R 2 values of the fitted curves were 0.9683 for MV3 cells and 0.9820 for B16F10 cells.

Article Snippet: Mouse melanoma B16F10 cell line, obtained from the American Type Culture Collection (ATCC), was maintained in DMEM (10-013-CV, Corning) with 10% FBS and 1% ( v / v ) penicillin–streptomycin.

Techniques: MTT Assay, Generated

Journal: International Journal of Molecular Sciences

Article Title: Disulfiram/Copper Combined with Irradiation Induces Immunogenic Cell Death in Melanoma

doi: 10.3390/ijms27020980

Figure Lengend Snippet: DSF/Cu (Cu: 1 μM) + IR induces apoptosis of MV3 and B16F10 melanoma cells. After MV3 and B16F10 cells were treated with DSF: 0–0.2 µM, Cu: 1 µM and IR (0, 12 Gy) for 24 h, apoptotic cell rate (%) is determined by measuring Annexin V/7-AAD expression using flow cytometry ( A , B ). n = 3, * p < 0.05; ** p < 0.01; ns represents no significant difference.

Article Snippet: Mouse melanoma B16F10 cell line, obtained from the American Type Culture Collection (ATCC), was maintained in DMEM (10-013-CV, Corning) with 10% FBS and 1% ( v / v ) penicillin–streptomycin.

Techniques: Expressing, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Disulfiram/Copper Combined with Irradiation Induces Immunogenic Cell Death in Melanoma

doi: 10.3390/ijms27020980

Figure Lengend Snippet: Cell surface detection of CRT by MV3 and B16F10 cells following treatment with DSF/Cu + IR. After treatment with (DSF: 0–0.2 µM, Cu: 1 µM) and IR (0, 12 Gy) for 24 h, Cell surface CRT expression of MV3 and B16F10 cells is determined by flow cytometry ( A , B ). n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Mouse melanoma B16F10 cell line, obtained from the American Type Culture Collection (ATCC), was maintained in DMEM (10-013-CV, Corning) with 10% FBS and 1% ( v / v ) penicillin–streptomycin.

Techniques: Expressing, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Disulfiram/Copper Combined with Irradiation Induces Immunogenic Cell Death in Melanoma

doi: 10.3390/ijms27020980

Figure Lengend Snippet: HMGB1 release from MV3 and B16F10 cells following treatment with DSF/Cu + IR. After treatment with DSF: 0–0.2 µM, Cu: 1 µM and IR (0, 12 Gy) for 24 h, HMGB1 release from MV3 and B16F10 cells was determined by ELISA ( A , B ). n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001; ns represents no significant difference.

Article Snippet: Mouse melanoma B16F10 cell line, obtained from the American Type Culture Collection (ATCC), was maintained in DMEM (10-013-CV, Corning) with 10% FBS and 1% ( v / v ) penicillin–streptomycin.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Disulfiram/Copper Combined with Irradiation Induces Immunogenic Cell Death in Melanoma

doi: 10.3390/ijms27020980

Figure Lengend Snippet: Intracellular ATP levels in MV3 and B16F10 cells following treatment with DSF/Cu + IR. After treatment with DSF: 0–0.2 µM, Cu: 1 µM and IR (0, 12 Gy) for 24 h, Intracellular ATP levels in MV3 and B16F10 cells were determined by flow cytometry ( A , B ). n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001; ns represents no significant difference.

Article Snippet: Mouse melanoma B16F10 cell line, obtained from the American Type Culture Collection (ATCC), was maintained in DMEM (10-013-CV, Corning) with 10% FBS and 1% ( v / v ) penicillin–streptomycin.

Techniques: Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Disulfiram/Copper Combined with Irradiation Induces Immunogenic Cell Death in Melanoma

doi: 10.3390/ijms27020980

Figure Lengend Snippet: DSF/Cu + IR inhibits growth of B16F10 melanoma in vivo. B16F10 cells were implanted in C57BL/6 mice. Untreated group: mice were left untreated. DSF/Cu group: mice received intratumoral (i.t.) injections of 100 µL PBS containing DSF/Cu (DSF, 0.3 µM; Cu, 1 µM) on days 1 and 3. IR (12 Gy, single dose) group: tumor-localized irradiation (12 Gy) was delivered after the first intratumoral DSF in-jection in each treatment cycle, for a total of three cycles, with the remainder of the body shielded by lead. DSF/Cu + IR group: mice received intratumoral (i.t.) injections of 100 µL PBS containing DSF/Cu (DSF, 0.3 µM; Cu, 1 µM) on days 1 and 3. Tumor-localized IR (12 Gy) was delivered once on day 2. This treatment cycle was repeated three times at 7-day intervals ( A ). Tumor growth was monitored every two days. Tumor growth ( B ), Photos of isolated tumors from each group ( C ) were displayed. n ≥ 5, ** p < 0.01; *** p < 0.001.

Article Snippet: Mouse melanoma B16F10 cell line, obtained from the American Type Culture Collection (ATCC), was maintained in DMEM (10-013-CV, Corning) with 10% FBS and 1% ( v / v ) penicillin–streptomycin.

Techniques: In Vivo, Irradiation, Isolation

Journal: International Journal of Molecular Sciences

Article Title: Disulfiram/Copper Combined with Irradiation Induces Immunogenic Cell Death in Melanoma

doi: 10.3390/ijms27020980

Figure Lengend Snippet: Flow cytometry analysis of tumor-infiltrating immune cells. Flow cytometry of tumor-infiltrating immune cells in B16F10 tumors following DSF/Cu and IR treatment. ( A ) The schematic of the gating strategy for CD8 + T cell, CD8 + T cell, DCs and MDSCs ( B – D ) The percentages of CD3 + T cells, CD3 + CD8 + T cells and CD3 + CD4 + T cells ( B ), DCs (CD11c + CD11b + cells ( C ) and MDSCs (CD11c + and GR-1 + cells) ( D ) in B16F10 tumors are displayed, respectively. No p value was reported because tumor cells from mice within each treatment group were pooled into a single sample, as tumors in the DSF/Cu + IR group were too small to analyze individually.

Article Snippet: Mouse melanoma B16F10 cell line, obtained from the American Type Culture Collection (ATCC), was maintained in DMEM (10-013-CV, Corning) with 10% FBS and 1% ( v / v ) penicillin–streptomycin.

Techniques: Flow Cytometry