atcc guidelines  (ATCC)

Journal: Frontiers in Immunology

Article Title: Docetaxel enhances Vβ-directed T-cell activation and antitumor immunity mediated by a bifunctional TCR agonist in breast and prostate cancer models

doi: 10.3389/fimmu.2026.1850760

Figure Lengend Snippet: Combination therapy with docetaxel and mSTAR1302 elicits antitumor effects in triple negative breast cancer (4T1) and androgen-independent prostate cancer (TRAMP-C2) mouse models. (A–E) 8–12-week-old female Balb/c mice were inoculated subcutaneously with 5x10 4 4T1 in the mammary fat pad (n=8–10 per group). (A) pictogram depicts experimental design. When the tumors reached 40-70mm 3 , mice were administered 3 doses of mSTAR1302 (1 mg/kg, i.p.) once per week on days 9, 16, and 23 and 3 doses of docetaxel (500 µg, i.p.), every other day, 7 days post-initial mSTAR1302 treatment, on days 16, 18, and 20. Primary tumors were measured in terms of mean (B) and individual (C) tumor volumes, as depicted. Inset numbers are tumor-free mice. In two additional cohorts, to assess metastatic growth, lungs were removed from 4T1 tumor-bearing mice 28 days post-tumor inoculation following the same treatment schedule. Lungs were dissociated, cultured in medium (2x FBS with 6-thioguanine) and incubated at 37 °C with 5% CO 2 for 12 days. (D) a meta-analysis from two separate studies is depicted with closed and opened circles differentiating data collected from each study. Cell colonies representing lung metastases were stained with 0.05% methylene blue and counted for each treatment group (n=14–20 per group total). Survival (E) was measured over time with inset numbers depicting median overall survival (mOS) and shaded bands depicting 95% confidence intervals. (F–I) 8–12-week-old male C57bl/6 mice were inoculated subcutaneously with 2.5x10 6 TRAMP-C2, on the right flank (n=8–10 per group). (F) pictogram depicts experimental design. Mice were administered mSTAR1302 (1 mg/kg, i.p.) when tumors reached 70–100mm 3 on days 22, 29, and 36, and docetaxel (500 µg, i.p.) on days 29, 31, and 33. Tumors were measured with mean (G) and individual (H) tumor volumes depicted. Inset numbers are tumor-free mice. Survival (I) was measured over time with mOS and 95% confidence intervals (shaded bands) depicted. Statistical tests: tumor growth: two-way ANOVA with Tukey’s post hoc test; comparison between groups: one-way ANOVA with Tukey’s post hoc test; survival; Mantel-Cox test. Error bars, SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. ANOVA, analysis of variance; FBS, fetal bovine serum; i.p., intraperitoneal; mets, metastases; s.c., subcutaneously.

Article Snippet: The murine TNBC cell line 4T1 and androgen-independent prostate cell line TRAMP-C2 were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA) and cultured according to ATCC recommendations.

Techniques: Cell Culture, Incubation, Staining, Comparison

Journal: Frontiers in Immunology

Article Title: Docetaxel enhances Vβ-directed T-cell activation and antitumor immunity mediated by a bifunctional TCR agonist in breast and prostate cancer models

doi: 10.3389/fimmu.2026.1850760

Figure Lengend Snippet: Docetaxel induces upregulation of death receptors TRAIL-R2 and FAS in 4T1 and TRAMP-C2 cells. (A) 4T1 and TRAMP-C2 cells were treated in vitro with either no drug or docetaxel (250 ng/mL) for 48 hours and analyzed for surface expression of FAS and TRAIL-R2 via flow cytometry. Histograms indicating frequency and gMFI are shown. Experiment repeated twice with similar results. (B) 4T1 and TRAMP-C2 cells were treated as described in Figure 1A and then co-cultured with TRAIL ligand or FAS ligand and protein G and measured for relative cell lysis compared to cells alone. (C, D) tumors harvested on day 28, after treatment as described in Figure 1A , were fixed for multiplex immunofluorescence staining and quantified for CD8 (green) and TRAIL-R2 or FAS (red) expression. (E) differential gene expression was interrogated using NanoString murine PanCancer immune profiling panel on tumors harvested on day 23 from 4T1 tumor-bearing Balb/c female mice, following treatment as previously described. Fold change compared to untreated tumors reported for TRAIL and FAS ligands. (F) 4T1 cells were exposed to docetaxel (250 ng/mL) for 48 hours and co-cultured with NK cells isolated from tumor-free Balb/c mice (E:T = 25:1) and monitored for relative NK-specific lysis of tumor cells for up to 8 hours post-effector addition. Statistical tests: comparison between groups: One-way ANOVA with Tukey’s post hoc test. comparison between two groups: unpaired T-test. Error bars, SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. ANOVA, analysis of variance; FoV, field of view; gMFI, geometric mean fluorescence intensity; NK, natural killer.

Article Snippet: The murine TNBC cell line 4T1 and androgen-independent prostate cell line TRAMP-C2 were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA) and cultured according to ATCC recommendations.

Techniques: In Vitro, Expressing, Flow Cytometry, Cell Culture, Lysis, Multiplex Assay, Immunofluorescence, Staining, Gene Expression, Isolation, Comparison, Fluorescence

Journal: Frontiers in Immunology

Article Title: Docetaxel enhances Vβ-directed T-cell activation and antitumor immunity mediated by a bifunctional TCR agonist in breast and prostate cancer models

doi: 10.3389/fimmu.2026.1850760

Figure Lengend Snippet: Antitumor activity of docetaxel and mSTAR1302 attenuated in TRAIL-R2 knockdown tumor mouse models. (A) TRAIL-R2 was knocked down via CRISPR in TRAMP-C2 and 4T1 cell lines. Flow cytometry was used to confirm expression in clones and WT cell lines. (B) 8–12-week-old male C57bl/6 mice were implanted with 2.5x10 6 TRAMP-C2 (n=10–12 per group) or TRAMP-C2 TRAIL-KO (n=7–9 per group) cells in the right flank. Treatment schedules for each group are depicted. Mean tumor volumes graphed. (C) 8–12-week-old female Balb/c mice were implanted with 5x10 4 4T1 (n=18–20 per group) or 4T1 TRAIL KO cells (n=10) in the mammary fat pad. WT 4T1 mice from this study were assessed concurrently in the study from Figure 7 and mean tumor volumes displayed in Figure 7C are the same tumor volumes displayed in Figure 7B . Statistical tests: tumor growth: two-way ANOVA with Tukey’s post hoc test. Error bars, SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. WT, wild type; KO, knockouts; CRISPR, clustered regularly interspaced short palindromic repeats; i.p., intraperitoneal; s.c., subcutaneously; ns, not significant.

Article Snippet: The murine TNBC cell line 4T1 and androgen-independent prostate cell line TRAMP-C2 were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA) and cultured according to ATCC recommendations.

Techniques: Activity Assay, Knockdown, CRISPR, Flow Cytometry, Expressing, Clone Assay

Journal: bioRxiv

Article Title: Aging-Associated Decline in Macrophage STAT6-OXPHOS Programs Promotes Tumor-Like Multinucleated Syncytia

doi: 10.64898/2026.05.18.726012

Figure Lengend Snippet: (A) Schematic illustration of STAT6 inhibition by AS1517499. IL-4R and IL-13R signaling activate STAT6-mediated transcriptional regulation, which was blocked using the STAT6 inhibitor AS1517499. (B) Experimental workflow for macrophage-tumor cell fusion assays. Tag-it Violet-labeled cancer cells (Py8119, mKRC.1, Py230, MyC-CaP, LLC1, and TRAMP-C2) were co-cultured with BMDMs or RAW264.7 macrophages in the presence of DMSO or AS1517499 for 72 h prior to flow cytometric and multiplex immunofluorescence analyses. (C) Flow cytometric quantification of TiV+ cells, TiV+/F4/80-cells, and TiV+/F4/80+ fused cells after treatment with DMSO or AS1517499. Data are presented as fold change relative to the DMSO control (n = 3 per group). (D) Quantification of TiV+/F4/80+ fused cells generated by co-culture of macrophages with different cancer cell lines (TRAMP-C2, Py8119, Py230, LLC1, MyC-CaP, and mKRC.1) in the presence of DMSO or AS1517499. Data are presented as fold change relative to the DMSO control (n ≥ 6 per group). (E) Representative immunofluorescence images of macrophages and multinucleated syncytia stained for F4/80, Ki67, and DAPI following DMSO or AS1517499 treatment. Yellow arrowheads indicate macrophage nuclei, and red arrowheads indicate Ki67-positive nuclei. Scale bar, 20 μm. (F) Quantification of the proportion of bi-nucleated (n = 40 in DMSO group and n = 39 in AS1517499 group) and multinucleated syncytia (n = 10 in DMSO group and n = 11 in AS1517499 group) containing Ki67-positive nuclei in DMSO- or AS1517499-treated co-cultures. Data are presented as mean ± SEM. Statistical significance was determined by the Mann-Whitney test. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 .

Article Snippet: L-929 (CCL-1), LLC1 (CRL-1642), MyC-CaP (CRL-3255), RAW264.7 (TIB-71), and TRAMP-C2 (CRL-2731) cells were obtained from ATCC.

Techniques: Inhibition, Labeling, Cell Culture, Multiplex Assay, Immunofluorescence, Control, Generated, Co-Culture Assay, Staining, MANN-WHITNEY

Journal: Biomolecules

Article Title: Induced Tumor-Suppressing (iTS) Cell-Based Approach for Protecting the Bone from Advanced Prostate Cancer

doi: 10.3390/biom16020240

Figure Lengend Snippet: Inhibitory effects of CM Oy-Lrp5 on TRAMP-C2ras prostate tumor cells. CN = control, CM = conditioned medium, Oy CM = osteocyte-derived CM, and Lrp5 = Lrp5 overexpression. The data are presented as mean ± S.D. (n = 3). The double asterisks indicate p < 0.01. ( a ) Inhibition of MTT-based viability of TRAMP-C2ras prostate tumor cells by CM Oy-Lrp5 . ( b ) Reduction in EdU-based proliferation by CM Oy-Lrp5 . Scale bar: 200 µm. ( c ) Increase in FRET efficiency in the vinculin biosensor by CM Oy-Lrp5 . The observed increase in FRET efficiency implies a decrease in vinculin-mediated molecular force at focal adhesions. Scale bar: 20 µm. ( d ) Inhibition of scratch-based migration of TRAMP-C2ras prostate tumor cells by CM Oy-Lrp5 . Scale bar: 200 µm. ( e ) Inhibition of transwell invasion by CM Oy-Lrp5 . Scale bar: 200 µm. ( f ) Reduction in the growth of 3D tumor spheroids by CM Oy-Lrp5 . Scale bar: 200 µm.

Article Snippet: TRAMP-C2ras prostate tumor cells (ATCC, CRL-2731) were cultured in DMEM/F-12 medium as described [ ].

Techniques: Control, Derivative Assay, Over Expression, Inhibition, Migration