bt474  (ATCC)

Journal: International Journal of Molecular Medicine

Article Title: Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2-positive breast cancer

doi: 10.3892/ijmm.2026.5751

Figure Lengend Snippet: EBA impairs cancer stem cell-like properties. (A) BT474 and SKBR3 cells were treated with EBA for 48 h, and ALDH1 activity was assessed by flow cytometry using the Aldefluor assay. DEAB was used to define the baseline of Aldefluor-positive fluorescence. (B) BT474 cells (5x10 4 cells/ml) were plated in ultra-low attachment dishes and cultured in the presence or absence of EBA for 5 days. The number and volume of mammospheres were measured by microscopy. (C) Overall survival of patients with breast cancer stratified by the co-expression of ALDH1A1 and CD44. (D) Spearman correlation analysis of ALDH1A1 and CD44 mRNA levels in patients with HER2-positive breast cancer from The Cancer Genome Atlas cohort (n=76). Kaplan-Meier survival analyses of patients with HER2-overexpressing breast cancer stratified by (E) ALDH1A1 and (F) CD44 expression. Patients were divided into high- and low-expression groups based on the median gene expression. Statistical significance was determined using the log-rank test. (G) JIMT-1 cells were treated with EBA (3 μ M) for 48 h and the CD44 high /CD24 low cell populations were identified by flow cytometry. (H) JIMT-1 cells (1.5x10 4 cells/ml) were cultured under serum-free suspension conditions in the presence of EBA (3 μ M) for 8 days. Mammosphere number and volumes were quantified. ** P<0.01 and **** P<0.0001 vs. vehicle-treated control (0 μ M EBA). EBA, ebastine; ALDH, aldehyde dehydrogenase; DEAB, diethylaminobenzaldehyde; CTL, control; ISO, isotype.

Article Snippet: The human breast cancer cell lines SKBR3, BT474, MDA-MB-453 (American Type Culture Collection) and JIMT-1 (Leibnitz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH) were cultured in DMEM, MEM or RPMI-1640 (all Sigma-Aldrich; Merck KGaA) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and 100 U/ml penicillin-streptomycin at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Activity Assay, Flow Cytometry, Fluorescence, Cell Culture, Microscopy, Expressing, Gene Expression, Suspension, Control

Journal: Journal of Advanced Research

Article Title: Single-cell and spatial transcriptome profiling identifies cellular heterogeneity and immunosuppressive tumor microenvironment in inflammatory breast cancer

doi: 10.1016/j.jare.2025.05.061

Figure Lengend Snippet: Tumor cell characteristics of IBC and non-IBC. A. Subpopulations of epithelial cells visualized using UMAP, revealing 13 major subpopulations across all analyzed samples, including luminal progenitor cells, mesenchymal cells, and epithelial proliferation cells, and epithelial1-10. B. Bar plots illustrating the proportion of each epithelial cell subtype within each type of samples. C. Representative enriched functional pathways based on DEGs of Epithelial 5 and Epithelial 1 between IBC1 and LBBC. D. Bar plots depicting the fraction of each epithelial cell subtype per sample. E. Representative hallmark pathways associated with marker genes of tumor clusters 1 and 4. F. Violin plot displaying the expression levels of HLA-A and FLNA in various tumor epithelial cell subpopulations, with higher expression observed in tumor epithelial cells from non-IBC patients compared to those from IBC patients. G. qRT-PCR analysis of HLA-A and FLNA expression in normal breast epithelial cells (MCF-10A) and various molecular subtype breast cancer cell lines (MCF-7, BT-474, MDA-MB-231, MDA-MB-453, SUM149PT, and KPL-4).

Article Snippet: MCF7, BT-474, MDA-MB-231, and MDA-MB-453 cell lines were purchased from the American Type Culture Collection (ATCC, Rockville, MD).

Techniques: Functional Assay, Marker, Expressing, Quantitative RT-PCR

Journal: bioRxiv

Article Title: A Developmental Lectin-Glycan Program Enables Early Breast Cancer Dissemination and Metastatic Onset

doi: 10.64898/2026.02.23.707584

Figure Lengend Snippet: A-B) LGALS1 mRNA (A) and protein (B) expression in JIMT-1 and BT-474 cell lines. C) GAL1 protein expression in JIMT-1 and BT-474 cell lines conditioned media (by ELISA). D) GAL1 binding on JIMT-1 and BT-474 cell lines (by flow cytometry). F) ST6GAL1 mRNA expression in JIMT-1 and BT-474 cell lines (qPCR). F) SNA binding on JIMT-1 and BT-474 cell lines (flow cytometry). G) Graphical abstract of analyzed cell lines summarizing their glycophenotype. H) Bright-field images of JIMT-1 cells treated with vehicle or rGAL1 (5μM, 72 h). I) Proliferation measured by tritiated thymidine incorporation after addition of rGAL1 (1, 3 or 5 µM) in JIMT-1 cells. J) Cell cycle analysis (PI+BrdU, flow cytometry). K) Quantification of the G1/ S /G2 phases. L) Heatmap of EMT and stemness markers (RT-qPCR, normalized). M) Dot plot of JIMT-1 cells treated with rGAL1 (5 µM) and stained for CD24 and CD44 (flow cytometry) (left panel), percentage of CD24 low /CD44 high cells across different experimental conditions (right panel). N) CD24 modulation normalized to mode, O) Mammosphere formation: cells pre-treated with rGal-1 (5 µM, 72 h), then plated in sphere media; spheres counted per 500 seeded cells. P) Wound-healing assay (Incucyte® 96-Well Woundmaker) Q) Wound confluency over 48hs. R) Graphical abstract of the tail vein injection experimental design. S) Lung images and quantification of macrometastasis at 6 weeks in KD or Scramble JIMT-1 injected-mice. Statistical analysis: (A-B-C-D-E-F-O-S) Unpaired Welch’s t-test. P-values < 0.05 were considered significant. (I-M) One-way ANOVA with Dunnet’s post-test (vs Vehicle). All experiments were performed with n ≥ 3 per group.

Article Snippet: BT-474 cell line was obtained from ATCC, while JIMT-1 cells were acquired from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Binding Assay, Flow Cytometry, Cell Cycle Assay, Quantitative RT-PCR, Staining, Wound Healing Assay, Injection