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: 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

Journal: Immunity

Article Title: Positioning and reversible suppression of CCR7 + dendritic cells in perivascular tumor niches shape cancer immunity

doi: 10.1016/j.immuni.2025.11.020

Figure Lengend Snippet: (A) Representative images and quantification of CCR7 + DCs (panCK − HLA-DR + LAMP3 + , yellow) near BVs (CD31 + PDPN − , magenta), or LVs (CD31 + PDPN + , cyan) in human tumors (HNSCC, NSCLC, and EC). Scale bar represents 20 μm. Whole-tumor sections were analyzed for EC and NSCLC. Numbers of fields of view (FOVs) analyzed per HNSCC sample are as follows: HNSCC1–04 n = 7; HNSCC1–06 n = 16; HNSCC1–07 n = 11; HNSCC2–01 n = 126; HNSCC2–06 n = 455; HNSCC2–09 n = 180; HNSCC2–11 n = 122; HNSCC2–12 n = 79; HNSCC2–15 n = 205; HNSCC2–26 n = 293; HNSCC2–35 n = 175. One bar = one patient . (B) Representative images and quantification of CCR7 + DCs (FSCN1 + ; yellow) located near BVs (CD31 + LYVE-1 − ; magenta) or LVs (CD31 + LYVE-1 + ; cyan) in mouse tumors (MC38, B16F10, and D4M3.A-OVA). Scale bar represents 10 μm. Whole-tumor sections were analyzed. One bar = one mouse. (C) Frequencies of BV-, LV- and non-vessel-associated CCR7 + DCs in mouse MC38 tumors 3 days post anti-CD40 or anti-PD-1 treatment. Whole-tumor sections were analyzed. One bar = one mouse. (D) (Left) Representative images of CCR7 + DCs (FSCN1 + ; yellow) located near BVs (CD31 + LYVE-1 − ; magenta) in MC38 tumors inoculated in Ccr7 ko/wt and Ccr7 ko/ko mice, 3 days post anti-PD-1 treatment. (Right) Distribution of the area of CCR7 + DC surfaces in clusters relative to their distance to closest BVs and plotted as percentage of total CCR7 + DC cluster area. CCR7 + DC surfaces from clusters associated with LVs and those not in clusters were excluded from the analysis. Scale bar represents 20 μm. Whole-tumor sections were analyzed. One dot = average value of all clusters in each genotype ( Ccr7 ko/ko n = 5 mice, 56 clusters; Ccr7 wt /ko n = 6 mice, 28 clusters; and Ccr7 wt /wt n = 3 mice, 19 clusters). Two-way ANOVA with multiple comparisons, mean with SEM; **** p < 0.0001 for comparison at 10 and 20 μm from closest BVs. (E) (Left) Representative images of CCR7 + DCs (FSCN1 + ; yellow) located near BVs (CD31 + LYVE-1 − ; magenta) and Ccl19 ( Ccl19 -eYFP + Tomato + ; white) in Ccl19 -ieYFP reporter mice (left image) or CCL21 (white, right image) in MC38 tumors. (Right) Frequencies of perivascular CCR7 + DC clusters associated with Ccl19 -covered BVs or within CCL21 + areas of the tumors among total perivascular CCR7 + DC clusters. Scale bar represents 20 μm. Whole-tumor sections were analyzed. One dot = one mouse. Unpaired t test, mean with SEM; *** p < 0.001. (F) (Left) Representative images of CCR7 + DCs (FSCN1 + ; yellow) located near BVs (CD31 + LYVE-1 − ; magenta) in MC38 tumors inoculated in Ccl19 wt/wt and Ccl19 ko/ko mice, 2 days post anti-PD-1treatment. (Right) Quantification of BV- or LV-associated CCR7 + DC clusters in MC38 tumors from Ccl19 wt/wt and Ccl19 ko/ko mice. Scale bar represents 20 μm. Whole-tumor sections were analyzed. One dot = one mouse, whiskers represent min to max. Unpaired t test; * p < 0.05. (G) Heatmap depicts log 2 -transformed averaged expression of Ccl19 in indicated immune and non-immune populations in the TME of multiple mouse tumor models (breast, , lung [and GSE201247 ], and pancreatic , ). (H) (Left) Synthetic images of CCR7 + DCs (yellow), blood endothelial cells (BECs; magenta), lymphatic endothelial cells (LECs; cyan), and CCL19 + fibroblasts (green) in one representative NSCLC patient analyzed by spatial transcriptomics. (Right) Box plots depict the enrichment scores of CCL19 + fibroblasts within the neighborhood of BV-associated CCR7 + DCs, in four human NSCLC. Data are shown for both permuted (median enrichment scores from 1,000 permutations) and observed datasets. Scale bar represents 20 μm. Whole-tumor sections were analyzed. One dot = one sample. Paired t test, whiskers represent mean to max; * p < 0.05. (I) Heatmap depicts log 2 -transformed averaged expression of CCL19 in indicated immune and non-immune populations in the TME of multiple human cancer types (HNSCC, n = 40, n = 18 patients; CRC, n = 23, n = 64 patients; ESCC, n = 58 patients ; NSCLC, n = 32, n = 7 patients; BRCA, n = 29 patients ; and PRCA, n = 18 patients ). A cross indicates that the cellular population was not detected. See also – .

Article Snippet: Murine B16F10 melanoma cell line , ATCC , CRL-6475; RRID: CVCL_0159.

Techniques: Comparison, Transformation Assay, Expressing, Spatial Transcriptomics