human breast cancer cell lines skbr3  (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: JID Innovations

Article Title: Human keratinocytes exhibit limited potential for SARS-CoV-2 infection despite ACE2 and mature cathepsin L expression

doi: 10.1016/j.xjidi.2025.100447

Figure Lengend Snippet: Quantification of ACE2 , dACE2 , TMPRSS2, NRP1, CTSL , BSG (CD147), DPP4 (CD26), AXL , KREMEN1 , and ASGR1 gene expression. mRNA levels normalized to GAPDH (2 -ΔCT ) in ( a ) NHDF, NHEKs, RHE, and human skin samples and in ( b ) Caco-2/TC-7, Calu-3, and HaCaT cell lines. Gene expression was quantified by RT-qPCR in triplicate. Data represent the mean ± SEM from ( a ) 3 to 14 independent experiments using biological samples from different individuals and ( b ) 3 or 4 independent experiments from cultured cells. NHDF, normal human dermal fibroblast; NHEK, normal human epidermal keratinocyte; RHE, reconstructed human epidermis.

Article Snippet: Human lung adenocarcinoma Calu-3 (Sterlab, Vallauris, France), human colon adenocarcinoma Caco-2/TC-7 (Sigma-Aldrich, St-Quentin-Fallavier, France), human epidermal keratinocyte cell line HaCaT (CLS, Köhln, Germany), and human promonocytic myeloid leukemia cell line U937 (ATCC, Manassas, VA) were maintained in complete MEM, IMDM, or DMEM medium (Gibco-Thermo Fisher Scientific, Courtaboeuf, France) supplemented with 10% fetal calf serum (Pierce, Thermo Fisher Scientific), penicillin (100 IU/ml), and streptomycin (100 μg/ml) (Gibco-Thermo Fisher Scientific) at 37 °C in a humidified atmosphere with 5% carbon dioxide.

Techniques: Gene Expression, Quantitative RT-PCR, Cell Culture

Journal: iScience

Article Title: Synthetic zipper mediated pre-targeting system for near-infrared photoimmunotherapy

doi: 10.1016/j.isci.2025.114558

Figure Lengend Snippet: The pre-targeting complex of NIR-PIT induces the release of ICD markers (A) Flow cytometric histograms show cell surface expression of calreticulin, HSP70, and HSP90 compared to untreated controls in SKOV3 cells with scFv-tisotumab-Zip2-Zip1-SNAP-IR700. (B) OVCAR3 cells treated with scFv-Sacituzumab-Zip2-Zip1-SNAP-IR700. (C) IGROV1 cells treated with scFv-Farletuzumab-Zip2-Zip1-SNAP-IR700. Only viable cells were included in the analysis. Bar graphs show the mean fluorescence intensity (MFI) of cell surface calreticulin, HSP70, and HSP90. Data are represented as mean ± SD from three biological replicates. Cells exposed to NIR light irradiation without incubation with NIR-PIT agents were used as control. Statistical significance was determined by a one-way ANOVA and Dunnett's test. ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001.

Article Snippet: Ovarian cancer cell lines SKOV3 (HTB-77), OVCAR3 (HTB-161), IGROV1 (SCC-203), A2780 (ECACC-93112519), and HEK293T (CRL-11268) were obtained from the American Type Culture Collection, the European Collection of Authenticated Cell Cultures, and Sigma-Aldrich.

Techniques: Expressing, Fluorescence, Irradiation, Incubation, Control

Journal: iScience

Article Title: Synthetic zipper mediated pre-targeting system for near-infrared photoimmunotherapy

doi: 10.1016/j.isci.2025.114558

Figure Lengend Snippet: The pre-targeting complex of NIR-PIT induces the release of ICD markers (A) Flow cytometric histograms show cell surface expression of calreticulin, HSP70, and HSP90 compared to untreated controls in SKOV3 cells with scFv-tisotumab-Zip2-Zip1-SNAP-IR700. (B) OVCAR3 cells treated with scFv-Sacituzumab-Zip2-Zip1-SNAP-IR700. (C) IGROV1 cells treated with scFv-Farletuzumab-Zip2-Zip1-SNAP-IR700. Only viable cells were included in the analysis. Bar graphs show the mean fluorescence intensity (MFI) of cell surface calreticulin, HSP70, and HSP90. Data are represented as mean ± SD from three biological replicates. Cells exposed to NIR light irradiation without incubation with NIR-PIT agents were used as control. Statistical significance was determined by a one-way ANOVA and Dunnett's test. ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001.

Article Snippet: Ovarian cancer cell lines SKOV3 (HTB-77), OVCAR3 (HTB-161), IGROV1 (SCC-203), A2780 (ECACC-93112519), and HEK293T (CRL-11268) were obtained from the American Type Culture Collection, the European Collection of Authenticated Cell Cultures, and Sigma-Aldrich.

Techniques: Expressing, Fluorescence, Irradiation, Incubation, Control

Journal: iScience

Article Title: Synthetic zipper mediated pre-targeting system for near-infrared photoimmunotherapy

doi: 10.1016/j.isci.2025.114558

Figure Lengend Snippet: The pre-targeting complex of NIR-PIT induces the release of ICD markers (A) Flow cytometric histograms show cell surface expression of calreticulin, HSP70, and HSP90 compared to untreated controls in SKOV3 cells with scFv-tisotumab-Zip2-Zip1-SNAP-IR700. (B) OVCAR3 cells treated with scFv-Sacituzumab-Zip2-Zip1-SNAP-IR700. (C) IGROV1 cells treated with scFv-Farletuzumab-Zip2-Zip1-SNAP-IR700. Only viable cells were included in the analysis. Bar graphs show the mean fluorescence intensity (MFI) of cell surface calreticulin, HSP70, and HSP90. Data are represented as mean ± SD from three biological replicates. Cells exposed to NIR light irradiation without incubation with NIR-PIT agents were used as control. Statistical significance was determined by a one-way ANOVA and Dunnett's test. ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001.

Article Snippet: SKOV3 , ATCC , HTB-77.

Techniques: Expressing, Fluorescence, Irradiation, Incubation, Control