murine tumor cell lines b16 melanoma  (ATCC)

Journal: Bioactive Materials

Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity

doi: 10.1016/j.bioactmat.2026.02.039

Figure Lengend Snippet: Physicochemical properties of BOLT, and BOLT reduces the growth of tumor cells. (A) Schematic of surface double-layer formation and ion release. (B) Negative zeta potential (−1.365 mV) and high conductivity (1.334 mS/cm), confirming colloidal stability and ion release. (C) Uniform particle size (∼1478 nm) across batches. (D) Interfacial pH buffering in PBS. (E) Naïve CD4 + T cells were isolated and activated using anti-CD3 and anti-CD28 using the culture media with 6.0 pH and treated with various doses of BOLT. RT-qPCR was performed to determine the expression of Gpcr68 at various BOLT doses in activated T cells at acidic pH. (F) Anti-CD3 and anti-CD28 activated CD4 + T cells were treated with different doses of BOLT to determine the protein expression of GPCR68 using Western blot. (G-J) CCK8 assay was performed to analyze the effect of various pH on B16, MC38, 143B, and MG63 cell proliferation. (K-L) Effect of various doses of BOLT on the B16 and K7M2 cell growth to determine the IC-50 of BOLT. Error bars represent mean ± SEM. ∗∗ p < 0.01 and ∗ p < 0.05.

Article Snippet: Murine tumor cell lines B16 melanoma (RRID: CVCL_0159), MC38 colon cancer (RRID: CVCL_B288), and 4T1 (RRID: CRL_2539) were purchased from the ATCC and cultured in RPMI 1640 medium (Gibco) or DMEM medium (Gibco) with 10% FBS as well as 1% penicillin/streptomycin.

Techniques: Zeta Potential Analyzer, Isolation, Quantitative RT-PCR, Expressing, Western Blot, CCK-8 Assay

Journal: Bioactive Materials

Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity

doi: 10.1016/j.bioactmat.2026.02.039

Figure Lengend Snippet: Anti-tumor effects of borate bioactive glass (BOLT) in B16 tumor. (A) Schematic illustration depicting the induction of B16 melanoma tumors, followed by treatment with BOLT at various time points, and tumor harvesting for subsequent analysis. (B) Tumor growth curves showing tumor volume in Control and BOLT-treated B16 melanoma tumors in mice. (C) Tumor weight at the time of harvesting in the BOLT-treated group compared to the Control. (D) Representative images of excised tumors from Control and BOLT-treated mice. (E) In vivo imaging of tumor-bearing mice in both the Control and BOLT-treated groups. (F) Flow cytometry analysis showing IFN-γ production in CD4 + and CD8 + T cells following BOLT treatment compared to Control. (G) Flow cytometry analysis demonstrated TNF-α production in CD4 + and CD8 + T cells in the BOLT-treated group, with a significant increase observed in CD8 + T cells. Student t-test was performed for comparison between the two groups. Two-way ANOVA was used for multiple comparisons. Data represent the mean ± SEM (n = 5). ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Murine tumor cell lines B16 melanoma (RRID: CVCL_0159), MC38 colon cancer (RRID: CVCL_B288), and 4T1 (RRID: CRL_2539) were purchased from the ATCC and cultured in RPMI 1640 medium (Gibco) or DMEM medium (Gibco) with 10% FBS as well as 1% penicillin/streptomycin.

Techniques: Control, In Vivo Imaging, Flow Cytometry, Comparison

Journal: Bioactive Materials

Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity

doi: 10.1016/j.bioactmat.2026.02.039

Figure Lengend Snippet: BOLT treatment induces ferroptosis in tumor cells. (A) RNA was extracted from Control and BOLT-treated tumors, and RNA sequencing (RNAseq) was performed to identify differentially expressed genes. (B) KEGG pathway analysis was conducted to assess the biological functions of the differentially expressed genes. (C) Heatmap displaying the differential expression of ferroptosis-related genes in BOLT-treated versus Control cells. (D) qRT-PCR analysis showing dose-dependent downregulation of Nrf2 in BOLT-treated cells. (E) qRT-PCR analysis of Duox1 expression in B16 cells following BOLT treatment. (F) Transmission electron microscopy (TEM) images showing mitochondrial shrinkage, increased membrane density, and loss of cristae in BOLT-treated cells. (G) Heatmap showing the dysregulated genes involved in ROS-chemical carcinogenesis in B16 cells treated with BOLT. (H) Flow cytometry analysis revealing reactive oxygen species (ROS) production in B16 cells treated with BOLT (0.25 μg/mL) compared to Control. (I) Histogram overlays and bar graph confirm elevated bodipy levels in BOLT-treated cells versus Control. (J) Annexin V/PI staining shows no significant apoptosis in B16 cells following BOLT treatment. (K) Western blot analysis showing the expression of genes involved in downregulating ferroptosis (SLC7A11, FACL4, and GPX4) in BOLT-treated B16 cells. Student t-test was performed for comparison between 2 groups. Two-way ANOVA was used for multiple comparisons. In-vitro experiments were performed in triplicate. Data are mean ± SEM, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Murine tumor cell lines B16 melanoma (RRID: CVCL_0159), MC38 colon cancer (RRID: CVCL_B288), and 4T1 (RRID: CRL_2539) were purchased from the ATCC and cultured in RPMI 1640 medium (Gibco) or DMEM medium (Gibco) with 10% FBS as well as 1% penicillin/streptomycin.

Techniques: Control, RNA Sequencing, RNA sequencing, Quantitative Proteomics, Quantitative RT-PCR, Expressing, Transmission Assay, Electron Microscopy, Membrane, Flow Cytometry, Staining, Western Blot, Comparison, In Vitro

Journal: Bioactive Materials

Article Title: pH-neutralization strategy to suppress GPCR68 spatiotemporally activates T cells and enhances anti-tumor immunity

doi: 10.1016/j.bioactmat.2026.02.039

Figure Lengend Snippet: Combinational treatment of BOLT and anti-CTLA-4 blockade enhances anti-tumor immune response in B16 melanoma. (A) C57BL/6 mice were subcutaneously injected with 1 × 10 5 B16 melanoma cells on day 0 to induce tumors. On day 7, mice were randomly divided into groups and treated with either BOLT alone (intratumoral injection administered on alternate days starting from day 7), anti-CTLA-4 (intraperitoneal injection administered on days 9, 11, 13, and 15), or a combination of both treatments. PBS was used as a vehicle Control, while IgG was used as anti-CTLA-4 Control. Tumor growth was monitored throughout the treatment period, and tumors were harvested for analysis on day 21. (B-C) Tumor growth curves and area under the curve (AUC) analysis for WT mice treated with BOLT, with or without anti-CTLA-4 antibody, following subcutaneous injection of B16 melanoma cells. Tumor growth was monitored, and analysis was conducted on day 21. (D) Representative images of excised tumors at day 21, showed reduced tumor size in combination-treated mice. (E, F) Flow cytometry analysis of IFN-γ production by tumor-infiltrating CD4 + and CD8 + T cells. (G, H) Flow cytometry analysis of TNF-α production by tumor-infiltrating CD4 + and CD8 + T cells. Two-way ANOVA was used for multiple comparisons. Data are mean ± SEM (n = 5), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Murine tumor cell lines B16 melanoma (RRID: CVCL_0159), MC38 colon cancer (RRID: CVCL_B288), and 4T1 (RRID: CRL_2539) were purchased from the ATCC and cultured in RPMI 1640 medium (Gibco) or DMEM medium (Gibco) with 10% FBS as well as 1% penicillin/streptomycin.

Techniques: Injection, Control, Flow Cytometry

Journal: Materials Today Bio

Article Title: Tetrahedral framework nucleic acids carrying tranexamic acid to alleviate ultraviolet B‐induced skin pigmentation

doi: 10.1016/j.mtbio.2026.103287

Figure Lengend Snippet: Assessment of the melanogenesis-inhibitory effects of tFNAs-TXA (A) Schematic workflow of the melanin inhibition assay. (B, C) CCK-8 assays showing the (B) cytotoxicity and (C) optimal concentration screening of tFNAs-TXA in B16 cells. (D) Morphological changes of B16 cells under different treatments. (E, F) EdU proliferation assay. Representative fluorescence images (E) and statistical analysis of EdU-positive rates (F) across the indicated groups (n = 6 independent biological replicates). Data are presented as mean ± SD. Statistical significance was assessed by one-way ANOVA test followed by Tukey's post-hoc test; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: Murine melanoma B16 cells were acquired from Procell Life Science & Technology Co., Ltd. (Wuhan, China) and cultured in RPMI-1640 medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) and penicillin-streptomycin (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C in a 5% CO 2 humidified atmosphere.

Techniques: Inhibition, CCK-8 Assay, Concentration Assay, Proliferation Assay, Fluorescence

Journal: Materials Today Bio

Article Title: Tetrahedral framework nucleic acids carrying tranexamic acid to alleviate ultraviolet B‐induced skin pigmentation

doi: 10.1016/j.mtbio.2026.103287

Figure Lengend Snippet: tFNAs-TXA inhibits melanin synthesis by suppressing the cAMP/CREB/MITF signaling pathway. (A) Representative immunofluorescence images showing the intracellular expression of MITF and TYR in B16 cells. Nuclei were stained with DAPI, cytoskeleton with Phalloidin, and target proteins with red fluorescence. (B) Quantitative analysis of relative fluorescence intensity for MITF and TYR (n = 6). (C) Representative Western blot bands showing the protein expression of CREB, p-CREB, MITF, and TYR (GAPDH served as the loading control). (D) Quantitative analysis of protein expression levels for Total CREB, p-CREB, MITF, and TYR, normalized to GAPDH. (E) Relative mRNA expression levels of Mitf and Tyr determined by RT-qPCR. Data are presented as mean ± SD (n = 6 independent biological replicates; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 by one-way ANOVA followed by Tukey's post-hoc test).

Article Snippet: Murine melanoma B16 cells were acquired from Procell Life Science & Technology Co., Ltd. (Wuhan, China) and cultured in RPMI-1640 medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) and penicillin-streptomycin (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C in a 5% CO 2 humidified atmosphere.

Techniques: Immunofluorescence, Expressing, Staining, Fluorescence, Western Blot, Control, Quantitative RT-PCR

Journal: bioRxiv

Article Title: ME1 Programs Latent Effector Capacity and Grounds a Mathematical Model of Reversible T Cell Exhaustion

doi: 10.64898/2026.05.05.722814

Figure Lengend Snippet: (A) Schematic illustrating the generation of CD8-specific ME1 transgenic (ME1 Tg) mice. (B) Representative flow cytometry showing tdTomato expression as a surrogate for ME1 overexpression in CD8 + tumor-infiltrating lymphocytes (TILs). (C–D) Average growth curves of B16F10 tumors in control and CD8-ME1 Tg mice treated with PBS (C) or combined anti-PD-1/anti-PD-L1 antibodies (100 μg each per dose) administered every other day starting on day 6 (arrow) after tumor implantation. Tumor growth was analyzed by two-way ANOVA (D, n = 6, **P < 0.01). (E) Tumor sizes were measured at endpoint. Data were analyzed using an unpaired two-tailed t test (n = 4–8, ***P < 0.001). One of two independent experiments is shown. (F) Flow cytometric analysis of granzyme B (GZMB) protein expression in CD8 + TILs cells from B16F10 tumors in ME1 Tg and control mice on day 12. Data were analyzed using an unpaired two-tailed Student’s t test ( * P< 0.05; n = 5 mice per group).

Article Snippet: B16F10 murine melanoma cell line was purchased from ATCC (CRL-6475) and cultured using DMEM complete medium.

Techniques: Transgenic Assay, Flow Cytometry, Expressing, Over Expression, Control, Tumor Implantation, Two Tailed Test