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human breast cancer cell lines skbr3  (ATCC)


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    ATCC human breast cancer cell lines skbr3
    EBA impairs cancer stem cell-like properties. (A) BT474 and <t>SKBR3</t> 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.
    Human Breast Cancer Cell Lines Skbr3, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1169 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+breast+cancer+cell+lines+skbr3/SK-BR-3/pmc12871574-35-1-9
    Average 97 stars, based on 1169 article reviews
    human breast cancer cell lines skbr3 - by Bioz Stars, 2026-10
    97/100 stars

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    1) Product Images from "Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2-positive breast cancer"

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

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2026.5751

    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.
    Figure Legend 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.

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

    Related Articles

    Multiple Displacement Amplification:

    Article Title: AIRE is expressed in breast cancer TANs and TAMs to regulate the extrinsic apoptotic pathway and inflammation.
    Article Snippet: .. Human breast cancer cell lines SKBR3, MDA-MB-231, MDA-MB-468, MCF-7, CAMA, and T47D; myeloid HL60 and THP-1; and mouse E0771 cells were all from the ATCC and cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (Biosera), 1% sodium pyruvate, 1% HEPES, and penicillin/ 666 | Journal of Leukocyte Biology, 2024, Vol. .. 115, No. 4 D ow nloaded from https://academ ic.oup.com /jleukbio/article/115/4/664/7462185 by guest on 07 D ecem ber 2024 streptomycin (100 U/mL and 100 μg/mL, respectively), except for the SKBR3 cell line that was cultured in McCoy’s 5 A medium (without phenol red) (Gibco) supplemented with 10% fetal bovine serum and penicillin/streptomycin.

    Article Title: Developing an antibody drug encapsulation nanoagent targeting HER2 for cancer treatment
    Article Snippet: .. Human breast cancer cell lines SKBR3, JIMT1, BT549, MDA-MB-231, and the human non-small cell lung cancer (NSCLC) cell line A549 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal bovine serum (FBS) and 1% Antibiotic-Antimycotic, at 37 °C and 5% CO2. .. DMEM and 10% FBS were purchased from Cytiva (Marlborough, MA, USA).

    Article Title: Developing an antibody drug encapsulation nanoagent targeting HER2 for cancer treatment.
    Article Snippet: .. Human breast cancer cell lines SKBR3, JIMT1, BT549, MDA-MB-231, and the human lung cancer cell line A549 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal bovine serum (FBS) and 1% Antibiotic-Antimycotic, at 37°C and 5% CO2. .. DMEM and 10% FBS were purchased from Cytiva (Marlborough, MA, USA).

    Article Title: Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2-positive breast cancer
    Article Snippet: Secondary antibodies included HRP-conjugated anti-mouse (Bio-Rad Laboratories, Inc.; cat. no. 1721011) and anti-rabbit IgG (Bio-Rad Laboratories, Inc.; cat. no. 1706515), as well as Alexa Fluor 594-conjugated goat anti-rabbit IgG (Invitrogen; Thermo Fisher Scientific, Inc.; cat. no. A-11037), Alexa Fluor 488-conjugated goat anti-rabbit IgG (Invitrogen; Thermo Fisher Scientific, Inc.; cat. no. A-11008), Alexa Fluor 594-conjugated goat anti-mouse IgG (Invitrogen; Thermo Fisher Scientific, Inc.; cat. no. A-11032) and Alexa Fluor 488-conjugated goat anti-mouse IgG (Invitrogen; Thermo Fisher Scientific, Inc.; cat. no. A-11001). .. 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 . .. All cell lines were passaged for <6 months and were authenticated by short tandem repeat profiling performed by Macrogen, Inc.

    Cell Culture:

    Article Title: AIRE is expressed in breast cancer TANs and TAMs to regulate the extrinsic apoptotic pathway and inflammation.
    Article Snippet: .. Human breast cancer cell lines SKBR3, MDA-MB-231, MDA-MB-468, MCF-7, CAMA, and T47D; myeloid HL60 and THP-1; and mouse E0771 cells were all from the ATCC and cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (Biosera), 1% sodium pyruvate, 1% HEPES, and penicillin/ 666 | Journal of Leukocyte Biology, 2024, Vol. .. 115, No. 4 D ow nloaded from https://academ ic.oup.com /jleukbio/article/115/4/664/7462185 by guest on 07 D ecem ber 2024 streptomycin (100 U/mL and 100 μg/mL, respectively), except for the SKBR3 cell line that was cultured in McCoy’s 5 A medium (without phenol red) (Gibco) supplemented with 10% fetal bovine serum and penicillin/streptomycin.

    Article Title: Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2‑positive breast cancer.
    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% CO2. .. All cell lines were passaged for <6 months and were authenticated by short tandem repeat profiling performed by Macrogen, Inc.

    Article Title: Developing an antibody drug encapsulation nanoagent targeting HER2 for cancer treatment
    Article Snippet: .. Human breast cancer cell lines SKBR3, JIMT1, BT549, MDA-MB-231, and the human non-small cell lung cancer (NSCLC) cell line A549 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal bovine serum (FBS) and 1% Antibiotic-Antimycotic, at 37 °C and 5% CO2. .. DMEM and 10% FBS were purchased from Cytiva (Marlborough, MA, USA).

    Article Title: Developing an antibody drug encapsulation nanoagent targeting HER2 for cancer treatment.
    Article Snippet: .. Human breast cancer cell lines SKBR3, JIMT1, BT549, MDA-MB-231, and the human lung cancer cell line A549 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal bovine serum (FBS) and 1% Antibiotic-Antimycotic, at 37°C and 5% CO2. .. DMEM and 10% FBS were purchased from Cytiva (Marlborough, MA, USA).

    Article Title: Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2-positive breast cancer
    Article Snippet: Secondary antibodies included HRP-conjugated anti-mouse (Bio-Rad Laboratories, Inc.; cat. no. 1721011) and anti-rabbit IgG (Bio-Rad Laboratories, Inc.; cat. no. 1706515), as well as Alexa Fluor 594-conjugated goat anti-rabbit IgG (Invitrogen; Thermo Fisher Scientific, Inc.; cat. no. A-11037), Alexa Fluor 488-conjugated goat anti-rabbit IgG (Invitrogen; Thermo Fisher Scientific, Inc.; cat. no. A-11008), Alexa Fluor 594-conjugated goat anti-mouse IgG (Invitrogen; Thermo Fisher Scientific, Inc.; cat. no. A-11032) and Alexa Fluor 488-conjugated goat anti-mouse IgG (Invitrogen; Thermo Fisher Scientific, Inc.; cat. no. A-11001). .. 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 . .. All cell lines were passaged for <6 months and were authenticated by short tandem repeat profiling performed by Macrogen, Inc.

    Modification:

    Article Title: Developing an antibody drug encapsulation nanoagent targeting HER2 for cancer treatment
    Article Snippet: .. Human breast cancer cell lines SKBR3, JIMT1, BT549, MDA-MB-231, and the human non-small cell lung cancer (NSCLC) cell line A549 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal bovine serum (FBS) and 1% Antibiotic-Antimycotic, at 37 °C and 5% CO2. .. DMEM and 10% FBS were purchased from Cytiva (Marlborough, MA, USA).

    Article Title: Developing an antibody drug encapsulation nanoagent targeting HER2 for cancer treatment.
    Article Snippet: .. Human breast cancer cell lines SKBR3, JIMT1, BT549, MDA-MB-231, and the human lung cancer cell line A549 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% Fetal bovine serum (FBS) and 1% Antibiotic-Antimycotic, at 37°C and 5% CO2. .. DMEM and 10% FBS were purchased from Cytiva (Marlborough, MA, USA).

    other:

    Article Title: Evaluation of nanobodies engineered with extra-cysteines for site-specific functionalization.
    Article Snippet: SKBR3 cells were cultured in McCoy’s 5 A medium (16600082, Gibco) supplemented with 10% fetal bovine serum (FBS, A5670201, Gibco) and 100 U/mL penicillin-streptomycin solution (15140122, Gibco).



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    human breast cancer cell lines skbr3  (ATCC)
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    ATCC human breast cancer cell lines skbr3
    EBA impairs cancer stem cell-like properties. (A) BT474 and <t>SKBR3</t> 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.
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    EBA impairs cancer stem cell-like properties. (A) BT474 and <t>SKBR3</t> 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.
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    SL-II CD16 + neutrophils retain tumor cell killing capacity but are deficient in immunosuppressive capacity (A) Cartoon of antibody dependent cellular cytotoxicity (ADCC) and myeloid derived suppressor cell (MDSC) activity exerted by mature activated neutrophils (Figure adapted from Aarts et al. ). (B) In vitro ADCC of LAN-1 cells unopsonized and opsonized (+Dinutuximab) ( n = 4) and <t>SKBR3</t> cells unopsonized and opsonized (+Trastuzumab) by PMNs (blue) ( n = 6) and SL-II CD16 + neutrophils (pink) ( n = 3) in a 1:50 T:E ratio. (C) Representative CFSE plots for CD8 + T cell proliferation of an in vitro MDSC activity assay. T cells were stimulated with anti-CD3/CD28 antibodies to induce proliferation and co-cultured with either unstimulated or TNFα-stimulated PMNs or SL-II CD16 + neutrophils. After 4 days, T cell proliferation was assessed by CFSE dilution ( n = 12 for T cells alone and n = 16 for co-culturing with PMNs or SL-II CD16 + neutrophils). (D) Boxplots of CD8 + T cell proliferation and damaged T cell formation in the in vitro MDSC assay. Unstimulated or TNFα stimulated PMNs (blue) ( n = 12) or SL-II CD16 + neutrophils (pink) ( n = 16) were co-cultured with T cells. After 4 days, FSC/SSC gating was used to assess ‘damaged’ T cell formation next to T cell proliferation. (E) Representative images taken with imaging flow cytometry of trogocytosis carried out by PMNs and SL-II CD16 + neutrophils. The scale bar was set at 10 μm ( n = 3). PMNs and SL-II CD16 + neutrophils were stained with calcein red-orange (orange), and T cells were stained with DiD (red) before co-culturing for 4 h. Data in (B) and (D) is represented as median and interquartile range. p values were calculated using Mann-Whitney U tests and labeled as ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.0001. n values represent the number of individual donor samples.
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    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.

    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

    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.

    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

    SL-II CD16 + neutrophils retain tumor cell killing capacity but are deficient in immunosuppressive capacity (A) Cartoon of antibody dependent cellular cytotoxicity (ADCC) and myeloid derived suppressor cell (MDSC) activity exerted by mature activated neutrophils (Figure adapted from Aarts et al. ). (B) In vitro ADCC of LAN-1 cells unopsonized and opsonized (+Dinutuximab) ( n = 4) and SKBR3 cells unopsonized and opsonized (+Trastuzumab) by PMNs (blue) ( n = 6) and SL-II CD16 + neutrophils (pink) ( n = 3) in a 1:50 T:E ratio. (C) Representative CFSE plots for CD8 + T cell proliferation of an in vitro MDSC activity assay. T cells were stimulated with anti-CD3/CD28 antibodies to induce proliferation and co-cultured with either unstimulated or TNFα-stimulated PMNs or SL-II CD16 + neutrophils. After 4 days, T cell proliferation was assessed by CFSE dilution ( n = 12 for T cells alone and n = 16 for co-culturing with PMNs or SL-II CD16 + neutrophils). (D) Boxplots of CD8 + T cell proliferation and damaged T cell formation in the in vitro MDSC assay. Unstimulated or TNFα stimulated PMNs (blue) ( n = 12) or SL-II CD16 + neutrophils (pink) ( n = 16) were co-cultured with T cells. After 4 days, FSC/SSC gating was used to assess ‘damaged’ T cell formation next to T cell proliferation. (E) Representative images taken with imaging flow cytometry of trogocytosis carried out by PMNs and SL-II CD16 + neutrophils. The scale bar was set at 10 μm ( n = 3). PMNs and SL-II CD16 + neutrophils were stained with calcein red-orange (orange), and T cells were stained with DiD (red) before co-culturing for 4 h. Data in (B) and (D) is represented as median and interquartile range. p values were calculated using Mann-Whitney U tests and labeled as ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.0001. n values represent the number of individual donor samples.

    Journal: iScience

    Article Title: Characterization of human CD34 + HSPC-derived neutrophils with limited myeloid-derived immunosuppressive cell activity

    doi: 10.1016/j.isci.2025.113404

    Figure Lengend Snippet: SL-II CD16 + neutrophils retain tumor cell killing capacity but are deficient in immunosuppressive capacity (A) Cartoon of antibody dependent cellular cytotoxicity (ADCC) and myeloid derived suppressor cell (MDSC) activity exerted by mature activated neutrophils (Figure adapted from Aarts et al. ). (B) In vitro ADCC of LAN-1 cells unopsonized and opsonized (+Dinutuximab) ( n = 4) and SKBR3 cells unopsonized and opsonized (+Trastuzumab) by PMNs (blue) ( n = 6) and SL-II CD16 + neutrophils (pink) ( n = 3) in a 1:50 T:E ratio. (C) Representative CFSE plots for CD8 + T cell proliferation of an in vitro MDSC activity assay. T cells were stimulated with anti-CD3/CD28 antibodies to induce proliferation and co-cultured with either unstimulated or TNFα-stimulated PMNs or SL-II CD16 + neutrophils. After 4 days, T cell proliferation was assessed by CFSE dilution ( n = 12 for T cells alone and n = 16 for co-culturing with PMNs or SL-II CD16 + neutrophils). (D) Boxplots of CD8 + T cell proliferation and damaged T cell formation in the in vitro MDSC assay. Unstimulated or TNFα stimulated PMNs (blue) ( n = 12) or SL-II CD16 + neutrophils (pink) ( n = 16) were co-cultured with T cells. After 4 days, FSC/SSC gating was used to assess ‘damaged’ T cell formation next to T cell proliferation. (E) Representative images taken with imaging flow cytometry of trogocytosis carried out by PMNs and SL-II CD16 + neutrophils. The scale bar was set at 10 μm ( n = 3). PMNs and SL-II CD16 + neutrophils were stained with calcein red-orange (orange), and T cells were stained with DiD (red) before co-culturing for 4 h. Data in (B) and (D) is represented as median and interquartile range. p values were calculated using Mann-Whitney U tests and labeled as ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.0001. n values represent the number of individual donor samples.

    Article Snippet: Human Breast Cancer Cell Line SKBR3 , ATCC , .

    Techniques: Derivative Assay, Activity Assay, In Vitro, Cell Culture, Imaging, Flow Cytometry, Staining, MANN-WHITNEY, Labeling