mouse melanoma cell line b16 f10  (ATCC)

Journal: MicrobiologyOpen

Article Title: Synergistic Effects of IMP‐1700, Ciprofloxacin, and X‐Ray Radiation in Bacteria and Mammalian Cell Lines: Implications for Use in Antimicrobial‐Resistant Bacteria

doi: 10.1002/mbo3.70270

Figure Lengend Snippet: Effect of 10 Gy X‐ray on viability of NIH‐3T3, B16.F10, MDA‐MB‐231, and HepG2 cells. Blue: untreated controls; Pink: irradiated cells. Cell counts were conducted at 24, 48, and 72 h post‐treatment. Statistical analysis was performed using two‐way ANOVA with Tukey's post hoc test: p < 0.05 (*), p < 0.005 (**), p < 0.0005 (***), p < 0.0001 (****), n = 3. Note: Y ‐axes are scaled independently for each cell line to account for differences in baseline growth, allowing clearer visualization of treatment effects. Tumor cell counts and representative images can be found in Appendix Table and Figure .

Article Snippet: The tumor cell lines, mouse melanoma B16.F10 (CRL‐6475, ATCC), human breast cancer MB‐MDA‐231 (CRM‐HTB‐26, ATCC), human hepatocellular carcinoma HepG2 C3A (a derivative of HepG2, CRL‐3581, ATCC), as well as the noncancerous mouse fibroblast cell line NIH‐3T3 (CRL‐1658, ATCC), were cultivated at 37°C, 5% CO2.

Techniques: Irradiation

Journal: MicrobiologyOpen

Article Title: Synergistic Effects of IMP‐1700, Ciprofloxacin, and X‐Ray Radiation in Bacteria and Mammalian Cell Lines: Implications for Use in Antimicrobial‐Resistant Bacteria

doi: 10.1002/mbo3.70270

Figure Lengend Snippet: Images of untreated NIH‐3T3, B16.F10, MDA‐MB‐231, and HepG2 cells and cells exposed to X‐ray. Scalebar: 300 µm.

Article Snippet: The tumor cell lines, mouse melanoma B16.F10 (CRL‐6475, ATCC), human breast cancer MB‐MDA‐231 (CRM‐HTB‐26, ATCC), human hepatocellular carcinoma HepG2 C3A (a derivative of HepG2, CRL‐3581, ATCC), as well as the noncancerous mouse fibroblast cell line NIH‐3T3 (CRL‐1658, ATCC), were cultivated at 37°C, 5% CO2.

Techniques:

Journal: MicrobiologyOpen

Article Title: Synergistic Effects of IMP‐1700, Ciprofloxacin, and X‐Ray Radiation in Bacteria and Mammalian Cell Lines: Implications for Use in Antimicrobial‐Resistant Bacteria

doi: 10.1002/mbo3.70270

Figure Lengend Snippet: Viability of NIH‐3T3, B16.F10, MDA‐MB‐231, and HepG2 cells after treatment with IMP‐1700 (5 µM), ciprofloxacin (CPX) (15 µM), their combination, or DMSO, with and without 10 Gy X‐ray. Cell counts were measured at 24, 48, and 72 h post‐treatment. Statistical significance assessed via two‐way ANOVA and Tukey's post hoc test: p < 0.05 (*), p < 0.005 (**), p < 0.0005 (***), p < 0.0001 (****), n = 3. Significant differences are only shown compared to untreated or 10 Gy. Tumor cell count values and images are available in Appendix Table and Figure , , , .

Article Snippet: The tumor cell lines, mouse melanoma B16.F10 (CRL‐6475, ATCC), human breast cancer MB‐MDA‐231 (CRM‐HTB‐26, ATCC), human hepatocellular carcinoma HepG2 C3A (a derivative of HepG2, CRL‐3581, ATCC), as well as the noncancerous mouse fibroblast cell line NIH‐3T3 (CRL‐1658, ATCC), were cultivated at 37°C, 5% CO2.

Techniques: Cell Characterization

Journal: MicrobiologyOpen

Article Title: Synergistic Effects of IMP‐1700, Ciprofloxacin, and X‐Ray Radiation in Bacteria and Mammalian Cell Lines: Implications for Use in Antimicrobial‐Resistant Bacteria

doi: 10.1002/mbo3.70270

Figure Lengend Snippet: Images of NIH‐3T3, B16.F10, MDA‐MB‐231, and HepG2 cells treated with IMP‐1700, with and without X‐ray. Scalebar: 300 µm.

Article Snippet: The tumor cell lines, mouse melanoma B16.F10 (CRL‐6475, ATCC), human breast cancer MB‐MDA‐231 (CRM‐HTB‐26, ATCC), human hepatocellular carcinoma HepG2 C3A (a derivative of HepG2, CRL‐3581, ATCC), as well as the noncancerous mouse fibroblast cell line NIH‐3T3 (CRL‐1658, ATCC), were cultivated at 37°C, 5% CO2.

Techniques:

Journal: MicrobiologyOpen

Article Title: Synergistic Effects of IMP‐1700, Ciprofloxacin, and X‐Ray Radiation in Bacteria and Mammalian Cell Lines: Implications for Use in Antimicrobial‐Resistant Bacteria

doi: 10.1002/mbo3.70270

Figure Lengend Snippet: Images of NIH‐3T3, B16.F10, MDA‐MB‐231, and HepG2 cells treated with ciprofloxacin (CPX), with and without X‐ray. Scalebar: 300 µm.

Article Snippet: The tumor cell lines, mouse melanoma B16.F10 (CRL‐6475, ATCC), human breast cancer MB‐MDA‐231 (CRM‐HTB‐26, ATCC), human hepatocellular carcinoma HepG2 C3A (a derivative of HepG2, CRL‐3581, ATCC), as well as the noncancerous mouse fibroblast cell line NIH‐3T3 (CRL‐1658, ATCC), were cultivated at 37°C, 5% CO2.

Techniques:

Journal: MicrobiologyOpen

Article Title: Synergistic Effects of IMP‐1700, Ciprofloxacin, and X‐Ray Radiation in Bacteria and Mammalian Cell Lines: Implications for Use in Antimicrobial‐Resistant Bacteria

doi: 10.1002/mbo3.70270

Figure Lengend Snippet: Images of NIH‐3T3, B16.F10, MDA‐MB‐231, and HepG2 cells treated with IMP‐1700 and ciprofloxacin (CPX), with and without X‐ray. Scalebar: 300 µm.

Article Snippet: The tumor cell lines, mouse melanoma B16.F10 (CRL‐6475, ATCC), human breast cancer MB‐MDA‐231 (CRM‐HTB‐26, ATCC), human hepatocellular carcinoma HepG2 C3A (a derivative of HepG2, CRL‐3581, ATCC), as well as the noncancerous mouse fibroblast cell line NIH‐3T3 (CRL‐1658, ATCC), were cultivated at 37°C, 5% CO2.

Techniques:

Journal: MicrobiologyOpen

Article Title: Synergistic Effects of IMP‐1700, Ciprofloxacin, and X‐Ray Radiation in Bacteria and Mammalian Cell Lines: Implications for Use in Antimicrobial‐Resistant Bacteria

doi: 10.1002/mbo3.70270

Figure Lengend Snippet: Images of NIH‐3T3, B16.F10, MDA‐MB‐231, and HepG2 cells treated with DMSO, with and without X‐ray. Scalebar: 300 µm.

Article Snippet: The tumor cell lines, mouse melanoma B16.F10 (CRL‐6475, ATCC), human breast cancer MB‐MDA‐231 (CRM‐HTB‐26, ATCC), human hepatocellular carcinoma HepG2 C3A (a derivative of HepG2, CRL‐3581, ATCC), as well as the noncancerous mouse fibroblast cell line NIH‐3T3 (CRL‐1658, ATCC), were cultivated at 37°C, 5% CO2.

Techniques:

Journal: Frontiers in Immunology

Article Title: ATG5-mediated inducible autophagy sustains CAR-T cell durability under solid tumor stress

doi: 10.3389/fimmu.2026.1720544

Figure Lengend Snippet: In vitro cytotoxicity of ATG5 or LC3b OE mCAR-T cells under an immunosuppressive TME–mimicking condition. (A) Schematic overview of the in vitro long-term cytotoxicity assay. mCAR-T cells were co-cultured with B16F0-hCD19-mCherry tumor cells at various effector-to-target (E:T) ratios (0.5:1 to 4:1) for 96 h in the presence of TGF-β (5 ng/mL) to mimic an immunosuppressive TME. Tumor cell confluency was quantified based on mCherry fluorescence intensity using a CellCyte™ live-cell imaging system. (B) Cytotoxicity kinetics of each mCAR-T cell group over time across different E:T ratios. Data represent mean ± SEM (n = 3). Statistical analysis was performed using two-way ANOVA followed by Tukey’s post hoc test to compare the main effects among pMIG, ATG5 OE, and LC3b OE CAR-T cells within each E:T ratio. *p < 0.05; **p < 0.01; ****p < 0.0001.

Article Snippet: The B16F0 (mouse melanoma) cell line was purchased from the American Type Culture Collection (ATCC), and the Plat-E (ecotropic retroviral packaging) cell line was obtained from Cell Biolabs (San Diego, CA, USA).

Techniques: In Vitro, Cytotoxicity Assay, Cell Culture, Fluorescence, Live Cell Imaging

Journal: Frontiers in Immunology

Article Title: ATG5-mediated inducible autophagy sustains CAR-T cell durability under solid tumor stress

doi: 10.3389/fimmu.2026.1720544

Figure Lengend Snippet: ATG5 OE reduces oxidative stress while enhancing effector function of mCAR-T cells under TGF-β–mediated immunosuppressive conditions. (A) Schematic illustration of the experimental design for assessing oxidative stress and effector functional characteristics. B16F0-hCD19 target cells (1 × 10 5 cells) were co-cultured with mCAR-T cells at an effector-to-target (E:T) ratio of 1:1 in the presence of TGF-β (5 ng/mL) and IL-2 (100 IU/mL). Intracellular cytokine staining (ICS) assays were performed in separate wells supplemented with GolgiPlug™, whereas ROS measurements and phenotypic analyses were conducted in parallel wells without GolgiPlug™. After 24 h of co-culture, cells were harvested and subjected to flow cytometric analysis. (B–E) Assessment of intracellular and mitochondrial ROS in mCAR-T cells co-cultured with B16F0-hCD19 tumor cells in the presence of TGF-β (5 ng/mL). (B) Representative histograms of total cellular ROS measured by CellROX™ Deep Red staining. (C) Quantification of cellular ROS levels expressed as mean fluorescence intensity (MFI ×10³). (D) Representative histograms of mitochondrial superoxide measured by MitoSOX™ Red staining. (E) Quantification of mitochondrial ROS levels expressed as MFI (×10²). In panels (C, E) , gray dotted lines indicate the basal status without tumor co-culture. (F–I) Functional characterization of mCAR-T cells following co-culture with B16F0-hCD19 tumor cells under TGF-β–mediated immunosuppressive conditions. (F) Representative histograms of intracellular perforin staining. (G) Quantification of perforin expression expressed as MFI (×10³). (H) Representative flow cytometry plots showing IFN-γ and TNF-α expression in pMIG, ATG5 OE, and LC3b OE mCAR-T cells. (I) Quantification of TNF-α + IFN-γ + cells among OE mCAR-T cells. (J, K) Analysis of exhaustion marker expression. (J) Representative flow cytometry plots showing PD-1 and TIM-3 expression in OE mCAR-T cells. (K) Quantification of PD-1 + TIM-3 + cells among OE mCAR-T cells. Individual replicates are shown as dots, and bars indicate mean ± SEM (n = 3). Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. See also Supplementary Figure 2 .

Article Snippet: The B16F0 (mouse melanoma) cell line was purchased from the American Type Culture Collection (ATCC), and the Plat-E (ecotropic retroviral packaging) cell line was obtained from Cell Biolabs (San Diego, CA, USA).

Techniques: Functional Assay, Cell Culture, Staining, Co-Culture Assay, Fluorescence, Expressing, Flow Cytometry, Marker

Journal: Frontiers in Immunology

Article Title: ATG5-mediated inducible autophagy sustains CAR-T cell durability under solid tumor stress

doi: 10.3389/fimmu.2026.1720544

Figure Lengend Snippet: ATG5 OE mCAR-T cells exhibit enhanced in vivo antitumor efficacy with irradiation preconditioning. (A) Schematic overview of the in vivo experimental design. C57BL/6 mice were subcutaneously inoculated with B16F0-hCD19 tumor cells (3 × 10 5 cells). In the IR group, whole-body irradiation (5 Gy) was administered on day 6, followed by adoptive transfer of hCD19 mCAR-T cells (5–7.5 × 10 6 cells) on day 7. In the Non-IR group, mCAR-T cells were adoptively transferred without irradiation. Tumor size was monitored at the indicated time points. (B) Kaplan–Meier survival analysis. Survival curves of tumor-bearing mice treated with untransduced CD8 + T cells (UTD), pMIG control, or ATG5 OE mCAR-T cells under Non-IR (left, dashed lines) or IR (right, solid lines) conditions. A tumor size ≥ 225 mm² was used as a surrogate endpoint for survival analysis for humane reasons. Statistical significance was assessed using the log-rank (Mantel-Cox) test, followed by Holm–Šídák’s multiple comparisons test for group comparisons. (C) Tumor growth kinetics. Mean tumor growth curves under Non-IR (left) and IR (right) conditions. Data are presented as mean ± SEM. Statistical significance was determined using two-way ANOVA with Tukey’s post hoc test. (D) Individual tumor growth trajectories. Tumor growth curves for individual mice treated with UTD, pMIG, or ATG5 OE mCAR-T cells under Non-IR (upper panels) and IR (lower panels) conditions. Group sizes were as follows: Non-IR, n = 15, 10, and 11; IR, n = 16, 10, and 12 for UTD CD8 + T cells, pMIG, and ATG5 OE mCAR-T cells, respectively. (E) Comparison of tumor sizes at defined time points. Tumor sizes measured on day 15 (Non-IR) and day 17 (IR). Each dot represents an individual mouse, and horizontal lines indicate mean values. Data are presented as mean ± SEM. Group sizes were as follows: Non-IR, n = 12, 8, and 10; IR, n = 11, 10, and 12 for UTD CD8 + T cells, pMIG, and ATG5 OE mCAR-T cells, respectively. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. *p < 0.05, **p < 0.01, ***p < 0.001, *** p < 0.0001. .

Article Snippet: The B16F0 (mouse melanoma) cell line was purchased from the American Type Culture Collection (ATCC), and the Plat-E (ecotropic retroviral packaging) cell line was obtained from Cell Biolabs (San Diego, CA, USA).

Techniques: In Vivo, Irradiation, Adoptive Transfer Assay, Control, Comparison

Journal: Frontiers in Immunology

Article Title: ATG5-mediated inducible autophagy sustains CAR-T cell durability under solid tumor stress

doi: 10.3389/fimmu.2026.1720544

Figure Lengend Snippet: ATG5 OE enhances the functional activity of mCAR-T cells in vivo under IR conditions. (A) Schematic overview of the in vivo tumor model and TIL analysis. C57BL/6 mice were subcutaneously inoculated with B16F0-hCD19 tumor cells (3 × 10 5 cells). In the IR group, mice received IR (5 Gy) on day 11, followed by adoptive transfer of anti-hCD19 mCAR-T cells (5 × 10 6 cells, i.v.) on day 12. In the Non-IR group, mCAR-T cells were transferred without IR. Tumors were harvested on day 20 for TIL analysis. (B, C) Tumor infiltration of CAR-T cells under Non-IR and IR conditions. (B) Representative flow cytometry plots showing CD8α and Thy1.1 expression among TILs under Non-IR (upper) and IR (lower) conditions. (C) Quantification of CD8 + CAR-T cells among total TILs. (D, E) Cytokine-producing capacity of CD8 + OE mCAR-TILs. The gating strategy is shown in Supplementary Figure 3A . (D) Representative flow cytometry plots showing IFN-γ and TNF-α expression. (E) Quantification of TNF-α + IFN-γ + cells among CD8 + OE mCAR-TILs. (F, G) Degranulation capacity of CD8 + OE mCAR-TILs. (F) Representative histograms of CD107a expression (IgG shown as a control). (G) Quantification of CD107a + cells among CD8 + OE mCAR-TILs. (H, I) Expression of exhaustion markers on CD8 + OE mCAR-TILs. (H) Representative flow cytometry plots showing PD-1 and TIM-3 expression. (I) Quantification of PD-1 + TIM-3 + cells among CD8 + OE mCAR-TILs. (J, K) Proliferative status of CD8 + OE mCAR-TILs. (J) Representative histograms of Ki67 expression (IgG shown as a control). (K) Quantification of Ki67 + cells among CD8 + OE mCAR-TILs. Individual replicates are shown as dots, and bars indicate mean ± SEM. Open dots represent the Non-IR group, and filled dots represent the IR group. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. See also Supplementary Figure 3 .

Article Snippet: The B16F0 (mouse melanoma) cell line was purchased from the American Type Culture Collection (ATCC), and the Plat-E (ecotropic retroviral packaging) cell line was obtained from Cell Biolabs (San Diego, CA, USA).

Techniques: Functional Assay, Activity Assay, In Vivo, Adoptive Transfer Assay, Flow Cytometry, Expressing, Control