human breast cancer cell line bt 474  (ATCC)


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    ATCC human breast cancer cell line bt 474
    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Human Breast Cancer Cell Line Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human breast cancer cell line bt 474 - by Bioz Stars, 2023-11
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    1) Product Images from "Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody"

    Article Title: Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody

    Journal: mAbs

    doi: 10.1080/19420862.2018.1486946

    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Figure Legend Snippet: ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.

    Techniques Used: Activity Assay

    human breast cancer cell lines bt474  (ATCC)


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    ATCC human breast cancer cell lines bt474
    Cytotoxicity (IC 50 ) of compounds and ADC toward cancer cell lines.
    Human Breast Cancer Cell Lines Bt474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    human breast cancer cell lines bt474 - by Bioz Stars, 2023-11
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    1) Product Images from "A HER2-targeted antibody-novel DNA topoisomerase I inhibitor conjugate induces durable adaptive antitumor immunity by activating dendritic cells"

    Article Title: A HER2-targeted antibody-novel DNA topoisomerase I inhibitor conjugate induces durable adaptive antitumor immunity by activating dendritic cells

    Journal: mAbs

    doi: 10.1080/19420862.2023.2220466

    Cytotoxicity (IC 50 ) of compounds and ADC toward cancer cell lines.
    Figure Legend Snippet: Cytotoxicity (IC 50 ) of compounds and ADC toward cancer cell lines.

    Techniques Used: Inhibition

    human breast cancer cell line bt 474  (ATCC)


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    ATCC human breast cancer cell line bt 474
    (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing <t>BT-474</t> cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.
    Human Breast Cancer Cell Line Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human breast cancer cell line bt 474/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human breast cancer cell line bt 474 - by Bioz Stars, 2023-11
    86/100 stars

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    1) Product Images from "Lysine-Directed Site-Selective Bioconjugation for the Creation of Radioimmunoconjugates"

    Article Title: Lysine-Directed Site-Selective Bioconjugation for the Creation of Radioimmunoconjugates

    Journal: Bioconjugate chemistry

    doi: 10.1021/acs.bioconjchem.2c00354

    (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing BT-474 cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.
    Figure Legend Snippet: (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing BT-474 cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.

    Techniques Used: Fluorescence, FACS, Expressing, Labeling, Magnetic Beads, Two Tailed Test, Software

    (A) Representative PET scans collected 24, 48, 72, 96, 120, and 144 h after the intravenous administration of [89Z]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, or [89Zr]Zr-malDFO-pertuzumab [3.7 −3.9 MBq (100-105 μCi), 20-21 μg, in 100 μL of PBS] to athymic nude mice bearing subcutaneous HER2-expressing BT-474 human breast cancer xenografts (n = 5). The images on the left are coronal slices, and those on the right are maximum intensity projections (MIPs); (B) biodistribution data from the mice used for PET imaging collected after the terminal imaging timepoint at 144 h (n = 5).
    Figure Legend Snippet: (A) Representative PET scans collected 24, 48, 72, 96, 120, and 144 h after the intravenous administration of [89Z]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, or [89Zr]Zr-malDFO-pertuzumab [3.7 −3.9 MBq (100-105 μCi), 20-21 μg, in 100 μL of PBS] to athymic nude mice bearing subcutaneous HER2-expressing BT-474 human breast cancer xenografts (n = 5). The images on the left are coronal slices, and those on the right are maximum intensity projections (MIPs); (B) biodistribution data from the mice used for PET imaging collected after the terminal imaging timepoint at 144 h (n = 5).

    Techniques Used: Expressing, Imaging

    human breast cancer cell line bt 474  (ATCC)


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    ATCC human breast cancer cell line bt 474
    (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing <t>BT-474</t> cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.
    Human Breast Cancer Cell Line Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human breast cancer cell line bt 474/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human breast cancer cell line bt 474 - by Bioz Stars, 2023-11
    86/100 stars

    Images

    1) Product Images from "Lysine-Directed Site-Selective Bioconjugation for the Creation of Radioimmunoconjugates"

    Article Title: Lysine-Directed Site-Selective Bioconjugation for the Creation of Radioimmunoconjugates

    Journal: Bioconjugate chemistry

    doi: 10.1021/acs.bioconjchem.2c00354

    (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing BT-474 cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.
    Figure Legend Snippet: (A) Fluorescence-associated cell sorting (FACS) of pertuzumab, DFO-pertuzumab, SSKDFO-pertuzumab, malDFO-pertuzumab, and non-specific hIgG1 using HER2-expressing BT-474 cells and an AlexaFluor488-labeled secondary antibody (n = 3); (B) bead-based immunoreactivity of [89Zr]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, and [89Zr]Zr-malDFO-pertuzumab using HER2-coated magnetic beads (n = 3). Statistical significance was determined via a two-tailed t test with a Welch’s correction using GraphPad Prism software. * = p-value < 0.05; ** = p-value < 0.01.

    Techniques Used: Fluorescence, FACS, Expressing, Labeling, Magnetic Beads, Two Tailed Test, Software

    (A) Representative PET scans collected 24, 48, 72, 96, 120, and 144 h after the intravenous administration of [89Z]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, or [89Zr]Zr-malDFO-pertuzumab [3.7 −3.9 MBq (100-105 μCi), 20-21 μg, in 100 μL of PBS] to athymic nude mice bearing subcutaneous HER2-expressing BT-474 human breast cancer xenografts (n = 5). The images on the left are coronal slices, and those on the right are maximum intensity projections (MIPs); (B) biodistribution data from the mice used for PET imaging collected after the terminal imaging timepoint at 144 h (n = 5).
    Figure Legend Snippet: (A) Representative PET scans collected 24, 48, 72, 96, 120, and 144 h after the intravenous administration of [89Z]Zr-DFO-pertuzumab, [89Zr]Zr-SSKDFO-pertuzumab, or [89Zr]Zr-malDFO-pertuzumab [3.7 −3.9 MBq (100-105 μCi), 20-21 μg, in 100 μL of PBS] to athymic nude mice bearing subcutaneous HER2-expressing BT-474 human breast cancer xenografts (n = 5). The images on the left are coronal slices, and those on the right are maximum intensity projections (MIPs); (B) biodistribution data from the mice used for PET imaging collected after the terminal imaging timepoint at 144 h (n = 5).

    Techniques Used: Expressing, Imaging

    human breast cancer cell lines bt474  (ATCC)


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    ATCC human breast cancer cell lines bt474
    PFKFB3 could be enhanced by hyperglycemia and was correlated with the prognosis of BC patients. A Protein levels of PFKFB3 in benign breast tissues and invasive ductal carcinoma with or without diabetes were detected by IHC. The magnification of the photographs was 200. B The Kaplan–Meier plotter database and C GEO database (GSE61304) were utilized to compare the PFS and OS of breast cancer patients with different PFKFB3 expression levels. D The breast cancer cell lines <t>(BT474</t> and MCF-7) were cultured in 1640 mediums with different concentrations of glucose (5.5 mM, 15 mM, or 25 mM) and WB was performed to evaluate PFKFB3 expression level. *, P < 0.05; **, P < 0.01; ***, P < 0.001
    Human Breast Cancer Cell Lines Bt474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Hyperglycemia induces PFKFB3 overexpression and promotes malignant phenotype of breast cancer through RAS/MAPK activation"

    Article Title: Hyperglycemia induces PFKFB3 overexpression and promotes malignant phenotype of breast cancer through RAS/MAPK activation

    Journal: World Journal of Surgical Oncology

    doi: 10.1186/s12957-023-02990-2

    PFKFB3 could be enhanced by hyperglycemia and was correlated with the prognosis of BC patients. A Protein levels of PFKFB3 in benign breast tissues and invasive ductal carcinoma with or without diabetes were detected by IHC. The magnification of the photographs was 200. B The Kaplan–Meier plotter database and C GEO database (GSE61304) were utilized to compare the PFS and OS of breast cancer patients with different PFKFB3 expression levels. D The breast cancer cell lines (BT474 and MCF-7) were cultured in 1640 mediums with different concentrations of glucose (5.5 mM, 15 mM, or 25 mM) and WB was performed to evaluate PFKFB3 expression level. *, P < 0.05; **, P < 0.01; ***, P < 0.001
    Figure Legend Snippet: PFKFB3 could be enhanced by hyperglycemia and was correlated with the prognosis of BC patients. A Protein levels of PFKFB3 in benign breast tissues and invasive ductal carcinoma with or without diabetes were detected by IHC. The magnification of the photographs was 200. B The Kaplan–Meier plotter database and C GEO database (GSE61304) were utilized to compare the PFS and OS of breast cancer patients with different PFKFB3 expression levels. D The breast cancer cell lines (BT474 and MCF-7) were cultured in 1640 mediums with different concentrations of glucose (5.5 mM, 15 mM, or 25 mM) and WB was performed to evaluate PFKFB3 expression level. *, P < 0.05; **, P < 0.01; ***, P < 0.001

    Techniques Used: Expressing, Cell Culture

    PFKFB3 might activate epithelial–mesenchymal transition and RAS/MAPK pathways of breast cancer in a hyperglycemic environment. A GSEA (Gene Set Enrichment Analysis) was performed to investigate the downstream signaling pathways. Pathway lists of PFKFB3 screened out by GSEA were shown on the left, the box plots of EMT (upper) and RAS/MAPK pathway (lower) were shown on the right. B Protein levels of PFKFB3, E-cadherin, N-cadherin, Vimentin, p-ERK1/2, and t-ERK1/2 in BT474-25 mM or MCF-7-25 mM cells after transfected with siPFKFB3-1, siPFKFB3-2 or siNC were detected using western blot. β-actin was detected as control. *, P < 0.05; **, P < 0.01
    Figure Legend Snippet: PFKFB3 might activate epithelial–mesenchymal transition and RAS/MAPK pathways of breast cancer in a hyperglycemic environment. A GSEA (Gene Set Enrichment Analysis) was performed to investigate the downstream signaling pathways. Pathway lists of PFKFB3 screened out by GSEA were shown on the left, the box plots of EMT (upper) and RAS/MAPK pathway (lower) were shown on the right. B Protein levels of PFKFB3, E-cadherin, N-cadherin, Vimentin, p-ERK1/2, and t-ERK1/2 in BT474-25 mM or MCF-7-25 mM cells after transfected with siPFKFB3-1, siPFKFB3-2 or siNC were detected using western blot. β-actin was detected as control. *, P < 0.05; **, P < 0.01

    Techniques Used: Transfection, Western Blot

    PFKFB3 promoted proliferation and migration of breast cancer cells in a hyperglycemic environment. BT474-25 mM and MCF-7-25 mM cells were transfected with siPFKFB3-1, siPFKFB3-2, or siNC. A The cell counting assay. B MTT assay. C The cell colony formation assay. D The wound-healing assay. E The migration assay
    Figure Legend Snippet: PFKFB3 promoted proliferation and migration of breast cancer cells in a hyperglycemic environment. BT474-25 mM and MCF-7-25 mM cells were transfected with siPFKFB3-1, siPFKFB3-2, or siNC. A The cell counting assay. B MTT assay. C The cell colony formation assay. D The wound-healing assay. E The migration assay

    Techniques Used: Migration, Transfection, Cell Counting, MTT Assay, Colony Assay, Wound Healing Assay

    Hyperglycemia might promote PFKFB3 expression by miR-26 downregulation in breast cancer. A TargetScan database, B OncomiR database, and C miRcode database were utilized to predict that miR-26 was a reliable upstream microRNA of PFKFB3. D Protein levels of PFKFB3, E-cadherin, N-cadherin, and Vimentin in BT474-25 mM or MCF-7-25 mM cells after transfected with miR-26-mimic, miR-26-inhibitor, or corresponding-NC were detected by western blot. β-actin was detected as control. *, P < 0.05; **, P < 0.01
    Figure Legend Snippet: Hyperglycemia might promote PFKFB3 expression by miR-26 downregulation in breast cancer. A TargetScan database, B OncomiR database, and C miRcode database were utilized to predict that miR-26 was a reliable upstream microRNA of PFKFB3. D Protein levels of PFKFB3, E-cadherin, N-cadherin, and Vimentin in BT474-25 mM or MCF-7-25 mM cells after transfected with miR-26-mimic, miR-26-inhibitor, or corresponding-NC were detected by western blot. β-actin was detected as control. *, P < 0.05; **, P < 0.01

    Techniques Used: Expressing, Transfection, Western Blot

    miR-26 downregulation accelerated the proliferation and migration of breast cancer in a hyperglycemic environment. BT474-25 mM and MCF-7-25 mM cells were transfected with miR-26-mimic, miR-26-inhibitor, or corresponding-NC. A The cell counting assay. B MTT assay. C The cell colony formation assay. D The wound-healing assay. E The migration assay. **, P < 0.01
    Figure Legend Snippet: miR-26 downregulation accelerated the proliferation and migration of breast cancer in a hyperglycemic environment. BT474-25 mM and MCF-7-25 mM cells were transfected with miR-26-mimic, miR-26-inhibitor, or corresponding-NC. A The cell counting assay. B MTT assay. C The cell colony formation assay. D The wound-healing assay. E The migration assay. **, P < 0.01

    Techniques Used: Migration, Transfection, Cell Counting, MTT Assay, Colony Assay, Wound Healing Assay

    human breast cancer cell lines bt474  (ATCC)


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    ATCC human breast cancer cell lines bt474
    Persistent activation of PI3K signaling promotes survival in lapatinib-resistant cells. (A) pHER2, total HER2, Akt T308 , Akt S473 , p70S6K S371 , 4EBP1 S65 , and survivin steady-state protein expression in untreated parental <t>BT474,</t> BT474 treated with 0.5 μ M lapatinib for 48 hours, and rBT474 maintained in 1 μ M lapatinib, as determined by Western blot analysis from whole cell extracts. (B) Phospho-PI3K protein expression was determined by RPMA in the same treatment groups as described in (A) . Results represent the mean ± standard error of triplicate samples, and are representative of three independent experiments. * P < 0.0018. (C) Molecular knockdown of PI3K by using pooled siRNA against PI3K subunits (*) in rBT474 cells was confirmed by Western blot analysis by using subunit-specific antibodies and an anti-PARP cleavage-product antibody. Cells transfected with scrambled siRNA construct (NSC) served as controls. Actin steady-state protein levels served as a control to ensure equal loading of protein. The results are representative of three independent experiments. (D) The effects of siRNA-mediated knockdown of PI3K on rBT474 cell growth ( P < 0.0058). Nonspecific siRNA construct (NSC) served as a control. Results represent the mean ± standard error of triplicate samples, and are representative of three independent experiments.
    Human Breast Cancer Cell Lines Bt474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human breast cancer cell lines bt474/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human breast cancer cell lines bt474 - by Bioz Stars, 2023-11
    86/100 stars

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    1) Product Images from "An heregulin-EGFR-HER3 autocrine signaling axis can mediate acquired lapatinib resistance in HER2+ breast cancer models"

    Article Title: An heregulin-EGFR-HER3 autocrine signaling axis can mediate acquired lapatinib resistance in HER2+ breast cancer models

    Journal: Breast Cancer Research : BCR

    doi: 10.1186/bcr3480

    Persistent activation of PI3K signaling promotes survival in lapatinib-resistant cells. (A) pHER2, total HER2, Akt T308 , Akt S473 , p70S6K S371 , 4EBP1 S65 , and survivin steady-state protein expression in untreated parental BT474, BT474 treated with 0.5 μ M lapatinib for 48 hours, and rBT474 maintained in 1 μ M lapatinib, as determined by Western blot analysis from whole cell extracts. (B) Phospho-PI3K protein expression was determined by RPMA in the same treatment groups as described in (A) . Results represent the mean ± standard error of triplicate samples, and are representative of three independent experiments. * P < 0.0018. (C) Molecular knockdown of PI3K by using pooled siRNA against PI3K subunits (*) in rBT474 cells was confirmed by Western blot analysis by using subunit-specific antibodies and an anti-PARP cleavage-product antibody. Cells transfected with scrambled siRNA construct (NSC) served as controls. Actin steady-state protein levels served as a control to ensure equal loading of protein. The results are representative of three independent experiments. (D) The effects of siRNA-mediated knockdown of PI3K on rBT474 cell growth ( P < 0.0058). Nonspecific siRNA construct (NSC) served as a control. Results represent the mean ± standard error of triplicate samples, and are representative of three independent experiments.
    Figure Legend Snippet: Persistent activation of PI3K signaling promotes survival in lapatinib-resistant cells. (A) pHER2, total HER2, Akt T308 , Akt S473 , p70S6K S371 , 4EBP1 S65 , and survivin steady-state protein expression in untreated parental BT474, BT474 treated with 0.5 μ M lapatinib for 48 hours, and rBT474 maintained in 1 μ M lapatinib, as determined by Western blot analysis from whole cell extracts. (B) Phospho-PI3K protein expression was determined by RPMA in the same treatment groups as described in (A) . Results represent the mean ± standard error of triplicate samples, and are representative of three independent experiments. * P < 0.0018. (C) Molecular knockdown of PI3K by using pooled siRNA against PI3K subunits (*) in rBT474 cells was confirmed by Western blot analysis by using subunit-specific antibodies and an anti-PARP cleavage-product antibody. Cells transfected with scrambled siRNA construct (NSC) served as controls. Actin steady-state protein levels served as a control to ensure equal loading of protein. The results are representative of three independent experiments. (D) The effects of siRNA-mediated knockdown of PI3K on rBT474 cell growth ( P < 0.0058). Nonspecific siRNA construct (NSC) served as a control. Results represent the mean ± standard error of triplicate samples, and are representative of three independent experiments.

    Techniques Used: Activation Assay, Expressing, Western Blot, Transfection, Construct

    Lapatinib-resistant cells exhibit a mixed pattern of EGFR tyrosine autophosphorylation. Reverse-phase protein microarray analysis of EGFR Y992, Y1068, Y1148, and Y1173 in parental HER2+ breast cancer cell lines (BT474; SKBR3; Au565; SUM190), parental cells treated with 1 μ M lapatinib for 24 hours, and lapatinib-resistant cell counterparts (rBT474; rSKBR3; rAu565; rSUM190) maintained in 1 μ M lapatinib. Results represent the mean ± standard error of triplicate samples and are representative of three independent experiments.
    Figure Legend Snippet: Lapatinib-resistant cells exhibit a mixed pattern of EGFR tyrosine autophosphorylation. Reverse-phase protein microarray analysis of EGFR Y992, Y1068, Y1148, and Y1173 in parental HER2+ breast cancer cell lines (BT474; SKBR3; Au565; SUM190), parental cells treated with 1 μ M lapatinib for 24 hours, and lapatinib-resistant cell counterparts (rBT474; rSKBR3; rAu565; rSUM190) maintained in 1 μ M lapatinib. Results represent the mean ± standard error of triplicate samples and are representative of three independent experiments.

    Techniques Used: Microarray

    Autocrine induction of HRG drives survival of resistant cells. (A) Immunofluorescence microscopy of HRG expression in the indicated cell lines by using a primary rabbit anti-HRG antibody and visualized with an anti-rabbit IgG Alexa Fluor 555 (red) conjugated secondary antibody. Cell nuclei were visualized by DAPI staining (blue). These results are representative of other fields on the slide. (B) Western blot analysis of HRG type 1 (115 kDa) and type 2 (40 kDa) steady-state protein levels in lapatinib-resistant (rSKBR3; rAu565; rBT474) maintained in 1 μ M lapatinib, and untreated parental cell counterparts (SKBR3; BT474; Au565); actin served as a control for equal loading of protein. (C) Western blot analysis of survivin and cleaved PARP product after HRG knockdown in rAu565 and rSKBR3 cells. Cells transfected with scrambled siRNA construct (NSC) served as controls. Actin served as a control for equal loading of protein. (D) Effects of siRNA-mediated knockdown of HRG on tumor cell growth in rAu565 and rSKBR3 cell lines. Results represent the mean ± standard error of triplicate samples, and are representative of three independent experiments. P < 0.009 (rSKBR3) and P < 0.0023 (rAu565). (E) Western blot analysis of ADAM17 protein level in parental BT474 and SKBR3 ± lapatinib treatment as indicated in the figure. Resistant cells (rBT474 and rSKBR3) were growing in the presence of 1 μ M lapatinib. Hela cell extract was used as a positive control. Two bands from 75 kDa to 100 kDa can be detected by a specific ADAM17 antibody. These results are representative of three independent experiments.
    Figure Legend Snippet: Autocrine induction of HRG drives survival of resistant cells. (A) Immunofluorescence microscopy of HRG expression in the indicated cell lines by using a primary rabbit anti-HRG antibody and visualized with an anti-rabbit IgG Alexa Fluor 555 (red) conjugated secondary antibody. Cell nuclei were visualized by DAPI staining (blue). These results are representative of other fields on the slide. (B) Western blot analysis of HRG type 1 (115 kDa) and type 2 (40 kDa) steady-state protein levels in lapatinib-resistant (rSKBR3; rAu565; rBT474) maintained in 1 μ M lapatinib, and untreated parental cell counterparts (SKBR3; BT474; Au565); actin served as a control for equal loading of protein. (C) Western blot analysis of survivin and cleaved PARP product after HRG knockdown in rAu565 and rSKBR3 cells. Cells transfected with scrambled siRNA construct (NSC) served as controls. Actin served as a control for equal loading of protein. (D) Effects of siRNA-mediated knockdown of HRG on tumor cell growth in rAu565 and rSKBR3 cell lines. Results represent the mean ± standard error of triplicate samples, and are representative of three independent experiments. P < 0.009 (rSKBR3) and P < 0.0023 (rAu565). (E) Western blot analysis of ADAM17 protein level in parental BT474 and SKBR3 ± lapatinib treatment as indicated in the figure. Resistant cells (rBT474 and rSKBR3) were growing in the presence of 1 μ M lapatinib. Hela cell extract was used as a positive control. Two bands from 75 kDa to 100 kDa can be detected by a specific ADAM17 antibody. These results are representative of three independent experiments.

    Techniques Used: Immunofluorescence, Microscopy, Expressing, Staining, Western Blot, Transfection, Construct, Positive Control

    human breast cancer cell line bt 474  (ATCC)


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    ATCC human breast cancer cell line bt 474
    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Human Breast Cancer Cell Line Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human breast cancer cell line bt 474/product/ATCC
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human breast cancer cell line bt 474 - by Bioz Stars, 2023-11
    94/100 stars

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    1) Product Images from "Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody"

    Article Title: Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody

    Journal: mAbs

    doi: 10.1080/19420862.2018.1486946

    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Figure Legend Snippet: ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.

    Techniques Used: Activity Assay

    human breast cancer cell lines bt 474  (ATCC)


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    ATCC human breast cancer cell lines bt 474
    Expression of HER2 in breast cancer cells, analyzed by flow cytometry. Notes: Representative fluorescence-activated cell sorting analysis of ( A ) MDA-MB-361 and ( B ) <t>BT-474</t> cells tested by ALDEFLUOR assay. In the right image (ALDH+ DEAB), cells incubated with ALDH substrate (BAAA) and the specific inhibitor of ALDH, DEAB, were used to establish the baseline fluorescence of these cells (R1) and to define the ALDEFLUOR-positive region (R2). ( C ) Percentage of HER2 positively stained cells in breast cancer cells. ( D ) HER2 mean fluorescence intensity in breast cancer cells. Data are expressed as mean ± SD (n=3). ** P <0.01. Abbreviations: ALDH, aldehyde dehydrogenase; BAAA, BODIPY-aminoacetaldehyde; DEAB, diethylaminobenzaldehyde.
    Human Breast Cancer Cell Lines Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells"

    Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S144184

    Expression of HER2 in breast cancer cells, analyzed by flow cytometry. Notes: Representative fluorescence-activated cell sorting analysis of ( A ) MDA-MB-361 and ( B ) BT-474 cells tested by ALDEFLUOR assay. In the right image (ALDH+ DEAB), cells incubated with ALDH substrate (BAAA) and the specific inhibitor of ALDH, DEAB, were used to establish the baseline fluorescence of these cells (R1) and to define the ALDEFLUOR-positive region (R2). ( C ) Percentage of HER2 positively stained cells in breast cancer cells. ( D ) HER2 mean fluorescence intensity in breast cancer cells. Data are expressed as mean ± SD (n=3). ** P <0.01. Abbreviations: ALDH, aldehyde dehydrogenase; BAAA, BODIPY-aminoacetaldehyde; DEAB, diethylaminobenzaldehyde.
    Figure Legend Snippet: Expression of HER2 in breast cancer cells, analyzed by flow cytometry. Notes: Representative fluorescence-activated cell sorting analysis of ( A ) MDA-MB-361 and ( B ) BT-474 cells tested by ALDEFLUOR assay. In the right image (ALDH+ DEAB), cells incubated with ALDH substrate (BAAA) and the specific inhibitor of ALDH, DEAB, were used to establish the baseline fluorescence of these cells (R1) and to define the ALDEFLUOR-positive region (R2). ( C ) Percentage of HER2 positively stained cells in breast cancer cells. ( D ) HER2 mean fluorescence intensity in breast cancer cells. Data are expressed as mean ± SD (n=3). ** P <0.01. Abbreviations: ALDH, aldehyde dehydrogenase; BAAA, BODIPY-aminoacetaldehyde; DEAB, diethylaminobenzaldehyde.

    Techniques Used: Expressing, Flow Cytometry, Fluorescence, FACS, Incubation, Staining

    Cell proliferation assay. Notes: ALDH+ and ALDH− breast cancer cells were seeded in 96-well plates with a density of 1×10 4 cells per well overnight. The cells were incubated with the nanoparticles or salinomycin for 72 h, and cell viability was evaluated using the CCK-8 assay. ( A ) MDA-MB-361 ALDH+, ( B ) MDA-MB-361 ALDH−, ( C ) BT-474 ALDH+, and ( D ) BT-474 ALDH−. Data are expressed as mean ± SD (n=3). Abbreviations: ALDH, aldehyde dehydrogenase; CCK-8, Cell Counting Kit-8; Sali-NP, salinomycin-loaded polymer-lipid nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.
    Figure Legend Snippet: Cell proliferation assay. Notes: ALDH+ and ALDH− breast cancer cells were seeded in 96-well plates with a density of 1×10 4 cells per well overnight. The cells were incubated with the nanoparticles or salinomycin for 72 h, and cell viability was evaluated using the CCK-8 assay. ( A ) MDA-MB-361 ALDH+, ( B ) MDA-MB-361 ALDH−, ( C ) BT-474 ALDH+, and ( D ) BT-474 ALDH−. Data are expressed as mean ± SD (n=3). Abbreviations: ALDH, aldehyde dehydrogenase; CCK-8, Cell Counting Kit-8; Sali-NP, salinomycin-loaded polymer-lipid nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Techniques Used: Proliferation Assay, Incubation, CCK-8 Assay, Cell Counting

    The IC 50 values of salinomycin and nanoparticles in breast cancer cells
    Figure Legend Snippet: The IC 50 values of salinomycin and nanoparticles in breast cancer cells

    Techniques Used:

    Effect of treatments on the percentage of CSCs. Notes: Effect of the treatments on the proportion of CSCs in breast cancer cells, as reflected by the tumorsphere formation rate ( A and B ) and the proportion of ALDH+ cells ( E and F ). Representative images of tumorspheres formed by ( C ) MDA-MB-361 cells and ( D ) BT-474 cells are shown. The rate of tumorsphere formation is defined as the number of tumorspheres formed in 7 days in the treatment group divided by the number of tumorspheres formed in 7 days in the untreated group; the rate of tumorsphere formation in the untreated group is used as a control and defined as 100%. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=6). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSCs, cancer stem cells; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.
    Figure Legend Snippet: Effect of treatments on the percentage of CSCs. Notes: Effect of the treatments on the proportion of CSCs in breast cancer cells, as reflected by the tumorsphere formation rate ( A and B ) and the proportion of ALDH+ cells ( E and F ). Representative images of tumorspheres formed by ( C ) MDA-MB-361 cells and ( D ) BT-474 cells are shown. The rate of tumorsphere formation is defined as the number of tumorspheres formed in 7 days in the treatment group divided by the number of tumorspheres formed in 7 days in the untreated group; the rate of tumorsphere formation in the untreated group is used as a control and defined as 100%. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=6). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSCs, cancer stem cells; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Techniques Used:

    In vivo antitumor assay in mice bearing subcutaneous BT-474 tumors. Notes: BT-474 tumors reached ~50 mm 3 in size by day 10. From day 10, mice were injected with nanoparticles (7.5 mg salinomycin/kg, i.v.) through the tail vein, and salinomycin (7.5 mg salinomycin/kg) dissolved in ethanol was administered by intraperitoneal injection. Therapy was given nine times on alternate days (indicated by arrows), and tumor volume was calculated. ( A ) Tumor growth curve. ( B ) Excised tumors. ( C ) The excised tumors were weighed at the end point. On day 28, the effect of the drug treatments on the CSC proportion in BT-474 tumors in vivo was evaluated by ( D ) the rate of tumorsphere formation and ( E ) the proportion of ALDH+ cells from the excised tumors. ( F ) Representative images of tumorspheres from ( D ) are shown. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=8). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSC, cancer stem cell; i.v., intravenous; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.
    Figure Legend Snippet: In vivo antitumor assay in mice bearing subcutaneous BT-474 tumors. Notes: BT-474 tumors reached ~50 mm 3 in size by day 10. From day 10, mice were injected with nanoparticles (7.5 mg salinomycin/kg, i.v.) through the tail vein, and salinomycin (7.5 mg salinomycin/kg) dissolved in ethanol was administered by intraperitoneal injection. Therapy was given nine times on alternate days (indicated by arrows), and tumor volume was calculated. ( A ) Tumor growth curve. ( B ) Excised tumors. ( C ) The excised tumors were weighed at the end point. On day 28, the effect of the drug treatments on the CSC proportion in BT-474 tumors in vivo was evaluated by ( D ) the rate of tumorsphere formation and ( E ) the proportion of ALDH+ cells from the excised tumors. ( F ) Representative images of tumorspheres from ( D ) are shown. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=8). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSC, cancer stem cell; i.v., intravenous; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Techniques Used: In Vivo, Injection

    human breast cancer cell lines bt474  (ATCC)


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    ATCC human breast cancer cell lines bt474
    (A) Expression of Darpp-32 (open bars) and t-Darpp (filled bars) mRNA in <t>BT474</t> cells and BT/Her R clones was analyzed by a SYBR Green assay. Shown are the average expression levels (±S.D.) of Darpp-32 and t-Darpp mRNAs in each indicated cell line relative to the corresponding mRNA levels in BT474 cells. Quadruplicate measurements were made on a single isolation of RNA from each cell line analyzed. Statistically significant differences from BT cells were determined by two-tailed t-test. *, p<0.05; **, p<0.01, ***, p<0.001. (B) Expression of Darpp-32 and t-Darpp proteins in BT474 and Her R /BT clones was analyzed by Western hybridization. SK-Br-3 cells exogenously expressing both t-Darpp and Darpp-32 (tDp/Dp32) were used as control for the respective forms of the protein; this lane was loaded with one-fifth as much lysate as the other lanes to account for their high exogenous t-Darpp/Darpp-32 expression. The top panel shows a short-exposure image of the membrane stained with an antibody (Santa Cruz H-62) that recognizes both Darpp-32 and t-Darpp. The middle panel is a long exposure of the same membrane and the bottom image shows the levels of actin as a loading control. Numbers below the actin image indicate the ratio of Darpp-32 to t-Darpp signal for each lane in the middle panel, as determined by densitometric scanning. Clones shown in this figure and additional BT/Her R clones have been analyzed multiple times by Western analysis, always with very high t-Darpp expression and low or undetectable expression of Darpp-32.
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    1) Product Images from "Darpp-32 and Its Truncated Variant t-Darpp Have Antagonistic Effects on Breast Cancer Cell Growth and Herceptin Resistance"

    Article Title: Darpp-32 and Its Truncated Variant t-Darpp Have Antagonistic Effects on Breast Cancer Cell Growth and Herceptin Resistance

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0006220

    (A) Expression of Darpp-32 (open bars) and t-Darpp (filled bars) mRNA in BT474 cells and BT/Her R clones was analyzed by a SYBR Green assay. Shown are the average expression levels (±S.D.) of Darpp-32 and t-Darpp mRNAs in each indicated cell line relative to the corresponding mRNA levels in BT474 cells. Quadruplicate measurements were made on a single isolation of RNA from each cell line analyzed. Statistically significant differences from BT cells were determined by two-tailed t-test. *, p<0.05; **, p<0.01, ***, p<0.001. (B) Expression of Darpp-32 and t-Darpp proteins in BT474 and Her R /BT clones was analyzed by Western hybridization. SK-Br-3 cells exogenously expressing both t-Darpp and Darpp-32 (tDp/Dp32) were used as control for the respective forms of the protein; this lane was loaded with one-fifth as much lysate as the other lanes to account for their high exogenous t-Darpp/Darpp-32 expression. The top panel shows a short-exposure image of the membrane stained with an antibody (Santa Cruz H-62) that recognizes both Darpp-32 and t-Darpp. The middle panel is a long exposure of the same membrane and the bottom image shows the levels of actin as a loading control. Numbers below the actin image indicate the ratio of Darpp-32 to t-Darpp signal for each lane in the middle panel, as determined by densitometric scanning. Clones shown in this figure and additional BT/Her R clones have been analyzed multiple times by Western analysis, always with very high t-Darpp expression and low or undetectable expression of Darpp-32.
    Figure Legend Snippet: (A) Expression of Darpp-32 (open bars) and t-Darpp (filled bars) mRNA in BT474 cells and BT/Her R clones was analyzed by a SYBR Green assay. Shown are the average expression levels (±S.D.) of Darpp-32 and t-Darpp mRNAs in each indicated cell line relative to the corresponding mRNA levels in BT474 cells. Quadruplicate measurements were made on a single isolation of RNA from each cell line analyzed. Statistically significant differences from BT cells were determined by two-tailed t-test. *, p<0.05; **, p<0.01, ***, p<0.001. (B) Expression of Darpp-32 and t-Darpp proteins in BT474 and Her R /BT clones was analyzed by Western hybridization. SK-Br-3 cells exogenously expressing both t-Darpp and Darpp-32 (tDp/Dp32) were used as control for the respective forms of the protein; this lane was loaded with one-fifth as much lysate as the other lanes to account for their high exogenous t-Darpp/Darpp-32 expression. The top panel shows a short-exposure image of the membrane stained with an antibody (Santa Cruz H-62) that recognizes both Darpp-32 and t-Darpp. The middle panel is a long exposure of the same membrane and the bottom image shows the levels of actin as a loading control. Numbers below the actin image indicate the ratio of Darpp-32 to t-Darpp signal for each lane in the middle panel, as determined by densitometric scanning. Clones shown in this figure and additional BT/Her R clones have been analyzed multiple times by Western analysis, always with very high t-Darpp expression and low or undetectable expression of Darpp-32.

    Techniques Used: Expressing, Clone Assay, SYBR Green Assay, Isolation, Two Tailed Test, Western Blot, Hybridization, Staining

    Several intracellular changes, including down-regulation of PKA-RIIα, PKIγ and PTG (green arrows) and up-regulation of t-Darpp (red arrow) work coordinately to enhance PKA activity. PKA, in turn, either activates the PI3K/Akt pathway (possibly through EGFR or the p85 regulatory subunit of PI3K) or promotes sustained phospho-Akt levels by activating the PP-1 inhibitory activity of Darpp-32. This activity is stimulated via a phosphorylation event at Thr-34, which is absent from t-Darpp. A second phosphorylation event, at Thr-75, activates Darpp-32 as a PKA inhibitor. We speculate that t-Darpp may interfere with this PKA inhibition via a dominant negative mechanism. Dashed lines indicate activities that are down-regulated in BT/Her R cells relative to BT474 cells. Question marks indicate hypothetical pathways that are activated in BT/Her R cells. All other pathways represent well established activities associated with the indicated proteins and enzymes. Abbreviations not already cited: PDK1/2, 3-phosphoinositide-dependent kinase-1/2; Pten, phosphatase and tensin homologue deleted in chromosome 10; PIP2, phosphatidylinositol-4,5-bisphosphate; PIP3, phosphatidylinositol-3,4,5-trisphosphate.
    Figure Legend Snippet: Several intracellular changes, including down-regulation of PKA-RIIα, PKIγ and PTG (green arrows) and up-regulation of t-Darpp (red arrow) work coordinately to enhance PKA activity. PKA, in turn, either activates the PI3K/Akt pathway (possibly through EGFR or the p85 regulatory subunit of PI3K) or promotes sustained phospho-Akt levels by activating the PP-1 inhibitory activity of Darpp-32. This activity is stimulated via a phosphorylation event at Thr-34, which is absent from t-Darpp. A second phosphorylation event, at Thr-75, activates Darpp-32 as a PKA inhibitor. We speculate that t-Darpp may interfere with this PKA inhibition via a dominant negative mechanism. Dashed lines indicate activities that are down-regulated in BT/Her R cells relative to BT474 cells. Question marks indicate hypothetical pathways that are activated in BT/Her R cells. All other pathways represent well established activities associated with the indicated proteins and enzymes. Abbreviations not already cited: PDK1/2, 3-phosphoinositide-dependent kinase-1/2; Pten, phosphatase and tensin homologue deleted in chromosome 10; PIP2, phosphatidylinositol-4,5-bisphosphate; PIP3, phosphatidylinositol-3,4,5-trisphosphate.

    Techniques Used: Activity Assay, Inhibition, Dominant Negative Mutation

    human breast cancer cell line bt474  (ATCC)


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    ATCC human breast cancer cell line bt474
    Trastuzumab resistant cells were developed by culturing parental <t>BT474</t> cells in the presence of 20/50 μ g trastuzumab for around 6 months.
    Human Breast Cancer Cell Line Bt474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "mRNA Profiling Reveals Determinants of Trastuzumab Efficiency in HER2-Positive Breast Cancer"

    Article Title: mRNA Profiling Reveals Determinants of Trastuzumab Efficiency in HER2-Positive Breast Cancer

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0117818

    Trastuzumab resistant cells were developed by culturing parental BT474 cells in the presence of 20/50 μ g trastuzumab for around 6 months.
    Figure Legend Snippet: Trastuzumab resistant cells were developed by culturing parental BT474 cells in the presence of 20/50 μ g trastuzumab for around 6 months.

    Techniques Used:

    (a) Proliferation rate of BT474 (parental) and trastuzumab resistant BT474 (BTR50) cells treated with 20 μ g/ml trastuzumab. Proliferation rates were measured daily over 7 days by a luciferase-based viability assay. (b) Sensitivity of BT474 (parental) and trastuzumab resistant BTR50 cells towards increasing concentrations of trastuzumab. Cell viability was determined by a luciferase-based viability assay after 7 days of treatment.
    Figure Legend Snippet: (a) Proliferation rate of BT474 (parental) and trastuzumab resistant BT474 (BTR50) cells treated with 20 μ g/ml trastuzumab. Proliferation rates were measured daily over 7 days by a luciferase-based viability assay. (b) Sensitivity of BT474 (parental) and trastuzumab resistant BTR50 cells towards increasing concentrations of trastuzumab. Cell viability was determined by a luciferase-based viability assay after 7 days of treatment.

    Techniques Used: Luciferase, Viability Assay

    The barchart displays the log2 fold changes of validated candidate genes, which significantly changed their expression in BT474 after trastuzumab treatment. The positive values indicate an upregulation upon drug treatment. Black bars denote values resulting from RNA-Seq analysis. Gray bars denote values resulting from RT-qPCR analysis.
    Figure Legend Snippet: The barchart displays the log2 fold changes of validated candidate genes, which significantly changed their expression in BT474 after trastuzumab treatment. The positive values indicate an upregulation upon drug treatment. Black bars denote values resulting from RNA-Seq analysis. Gray bars denote values resulting from RT-qPCR analysis.

    Techniques Used: Expressing, RNA Sequencing Assay, Quantitative RT-PCR

    Upregulated genes in  BT474  upon trastuzumab treatment.
    Figure Legend Snippet: Upregulated genes in BT474 upon trastuzumab treatment.

    Techniques Used: Binding Assay

    The barchart displays the log2 fold changes of validated candidate genes, which showed significant differences in their expression in BT474 and HCC1954, respectively. Positive values indicate an upregulation in BT474. Negative values indicate an upregulation in HCC1954. Black bars denote values resulting from RNA-Seq analysis. Gray bars denote values resulting from RT-qPCR analysis.
    Figure Legend Snippet: The barchart displays the log2 fold changes of validated candidate genes, which showed significant differences in their expression in BT474 and HCC1954, respectively. Positive values indicate an upregulation in BT474. Negative values indicate an upregulation in HCC1954. Black bars denote values resulting from RNA-Seq analysis. Gray bars denote values resulting from RT-qPCR analysis.

    Techniques Used: Expressing, RNA Sequencing Assay, Quantitative RT-PCR

    Differentially expressed genes between HCC1954 and  BT474.
    Figure Legend Snippet: Differentially expressed genes between HCC1954 and BT474.

    Techniques Used: Binding Assay

    The plot displays the result of a PCA on the pairwise normalized RNA-Seq expression values of all 23367 annotated genes in the samples BT474, BTR50 and HCC1954 with and without trastuzumab (T) treatment. Same colors denote that the samples belong to the same conducted statistical test and thus were normalized in pair. The five two sample tests included BT474 vs. HCC1954 , BT474 vs. BTR50 , HCC1954+T vs. HCC1954 , BTR50+T vs. BTR50 , and BT474+T vs. BT474 .
    Figure Legend Snippet: The plot displays the result of a PCA on the pairwise normalized RNA-Seq expression values of all 23367 annotated genes in the samples BT474, BTR50 and HCC1954 with and without trastuzumab (T) treatment. Same colors denote that the samples belong to the same conducted statistical test and thus were normalized in pair. The five two sample tests included BT474 vs. HCC1954 , BT474 vs. BTR50 , HCC1954+T vs. HCC1954 , BTR50+T vs. BTR50 , and BT474+T vs. BT474 .

    Techniques Used: RNA Sequencing Assay, Expressing

    SNPs called in the  BT474  cell line.
    Figure Legend Snippet: SNPs called in the BT474 cell line.

    Techniques Used:

    human bt474 breast cancer cell line  (ATCC)


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    ATCC human bt474 breast cancer cell line
    Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the <t>BT474</t> breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)
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    1) Product Images from "Phosphorylation of Ser 78 of Hsp27 correlated with HER-2/ neu status and lymph node positivity in breast cancer"

    Article Title: Phosphorylation of Ser 78 of Hsp27 correlated with HER-2/ neu status and lymph node positivity in breast cancer

    Journal: Molecular Cancer

    doi: 10.1186/1476-4598-6-52

    Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the BT474 breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)
    Figure Legend Snippet: Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the BT474 breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)

    Techniques Used: SDS Page, Cell Culture

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    ATCC human breast cancer cell line bt 474
    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.
    Human Breast Cancer Cell Line Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC human breast cancer cell lines bt474
    Cytotoxicity (IC 50 ) of compounds and ADC toward cancer cell lines.
    Human Breast Cancer Cell Lines Bt474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    ATCC human breast cancer cell lines bt 474
    Expression of HER2 in breast cancer cells, analyzed by flow cytometry. Notes: Representative fluorescence-activated cell sorting analysis of ( A ) MDA-MB-361 and ( B ) <t>BT-474</t> cells tested by ALDEFLUOR assay. In the right image (ALDH+ DEAB), cells incubated with ALDH substrate (BAAA) and the specific inhibitor of ALDH, DEAB, were used to establish the baseline fluorescence of these cells (R1) and to define the ALDEFLUOR-positive region (R2). ( C ) Percentage of HER2 positively stained cells in breast cancer cells. ( D ) HER2 mean fluorescence intensity in breast cancer cells. Data are expressed as mean ± SD (n=3). ** P <0.01. Abbreviations: ALDH, aldehyde dehydrogenase; BAAA, BODIPY-aminoacetaldehyde; DEAB, diethylaminobenzaldehyde.
    Human Breast Cancer Cell Lines Bt 474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human breast cancer cell lines bt 474/product/ATCC
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    human breast cancer cell lines bt 474 - by Bioz Stars, 2023-11
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    86
    ATCC human breast cancer cell line bt474
    Trastuzumab resistant cells were developed by culturing parental <t>BT474</t> cells in the presence of 20/50 μ g trastuzumab for around 6 months.
    Human Breast Cancer Cell Line Bt474, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human breast cancer cell line bt474/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human breast cancer cell line bt474 - by Bioz Stars, 2023-11
    86/100 stars
      Buy from Supplier

    95
    ATCC human bt474 breast cancer cell line
    Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the <t>BT474</t> breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)
    Human Bt474 Breast Cancer Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human bt474 breast cancer cell line/product/ATCC
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    Price from $9.99 to $1999.99
    human bt474 breast cancer cell line - by Bioz Stars, 2023-11
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    Image Search Results


    ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.

    Journal: mAbs

    Article Title: Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody

    doi: 10.1080/19420862.2018.1486946

    Figure Lengend Snippet: ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.

    Article Snippet: Human breast cancer cell line BT-474 (HTB-20), MDA-MB-175VII (HTB-25), SK-BR-3(HTB-30) and HCC1419(CRL-2326) are HER2-positive cell lines, which were obtained from the ATCC, and HCC1419 is trastuzumab-resistant human breast cancer cell line.

    Techniques: Activity Assay

    Cytotoxicity (IC 50 ) of compounds and ADC toward cancer cell lines.

    Journal: mAbs

    Article Title: A HER2-targeted antibody-novel DNA topoisomerase I inhibitor conjugate induces durable adaptive antitumor immunity by activating dendritic cells

    doi: 10.1080/19420862.2023.2220466

    Figure Lengend Snippet: Cytotoxicity (IC 50 ) of compounds and ADC toward cancer cell lines.

    Article Snippet: The following cells lines were purchased from the American Type Culture Collection: human breast cancer cell lines BT474 and SK-BR-3, human gastric cancer cell line NCI-N87, human pancreatic cancer cell line BxPC-3, mouse colon cancer cell line CT26.WT, and mouse breast cancer cell line EMT6.

    Techniques: Inhibition

    Expression of HER2 in breast cancer cells, analyzed by flow cytometry. Notes: Representative fluorescence-activated cell sorting analysis of ( A ) MDA-MB-361 and ( B ) BT-474 cells tested by ALDEFLUOR assay. In the right image (ALDH+ DEAB), cells incubated with ALDH substrate (BAAA) and the specific inhibitor of ALDH, DEAB, were used to establish the baseline fluorescence of these cells (R1) and to define the ALDEFLUOR-positive region (R2). ( C ) Percentage of HER2 positively stained cells in breast cancer cells. ( D ) HER2 mean fluorescence intensity in breast cancer cells. Data are expressed as mean ± SD (n=3). ** P <0.01. Abbreviations: ALDH, aldehyde dehydrogenase; BAAA, BODIPY-aminoacetaldehyde; DEAB, diethylaminobenzaldehyde.

    Journal: International Journal of Nanomedicine

    Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

    doi: 10.2147/IJN.S144184

    Figure Lengend Snippet: Expression of HER2 in breast cancer cells, analyzed by flow cytometry. Notes: Representative fluorescence-activated cell sorting analysis of ( A ) MDA-MB-361 and ( B ) BT-474 cells tested by ALDEFLUOR assay. In the right image (ALDH+ DEAB), cells incubated with ALDH substrate (BAAA) and the specific inhibitor of ALDH, DEAB, were used to establish the baseline fluorescence of these cells (R1) and to define the ALDEFLUOR-positive region (R2). ( C ) Percentage of HER2 positively stained cells in breast cancer cells. ( D ) HER2 mean fluorescence intensity in breast cancer cells. Data are expressed as mean ± SD (n=3). ** P <0.01. Abbreviations: ALDH, aldehyde dehydrogenase; BAAA, BODIPY-aminoacetaldehyde; DEAB, diethylaminobenzaldehyde.

    Article Snippet: Human breast cancer cell lines BT-474 and MDA-MB-361 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Expressing, Flow Cytometry, Fluorescence, FACS, Incubation, Staining

    Cell proliferation assay. Notes: ALDH+ and ALDH− breast cancer cells were seeded in 96-well plates with a density of 1×10 4 cells per well overnight. The cells were incubated with the nanoparticles or salinomycin for 72 h, and cell viability was evaluated using the CCK-8 assay. ( A ) MDA-MB-361 ALDH+, ( B ) MDA-MB-361 ALDH−, ( C ) BT-474 ALDH+, and ( D ) BT-474 ALDH−. Data are expressed as mean ± SD (n=3). Abbreviations: ALDH, aldehyde dehydrogenase; CCK-8, Cell Counting Kit-8; Sali-NP, salinomycin-loaded polymer-lipid nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Journal: International Journal of Nanomedicine

    Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

    doi: 10.2147/IJN.S144184

    Figure Lengend Snippet: Cell proliferation assay. Notes: ALDH+ and ALDH− breast cancer cells were seeded in 96-well plates with a density of 1×10 4 cells per well overnight. The cells were incubated with the nanoparticles or salinomycin for 72 h, and cell viability was evaluated using the CCK-8 assay. ( A ) MDA-MB-361 ALDH+, ( B ) MDA-MB-361 ALDH−, ( C ) BT-474 ALDH+, and ( D ) BT-474 ALDH−. Data are expressed as mean ± SD (n=3). Abbreviations: ALDH, aldehyde dehydrogenase; CCK-8, Cell Counting Kit-8; Sali-NP, salinomycin-loaded polymer-lipid nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Article Snippet: Human breast cancer cell lines BT-474 and MDA-MB-361 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Proliferation Assay, Incubation, CCK-8 Assay, Cell Counting

    The IC 50 values of salinomycin and nanoparticles in breast cancer cells

    Journal: International Journal of Nanomedicine

    Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

    doi: 10.2147/IJN.S144184

    Figure Lengend Snippet: The IC 50 values of salinomycin and nanoparticles in breast cancer cells

    Article Snippet: Human breast cancer cell lines BT-474 and MDA-MB-361 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques:

    Effect of treatments on the percentage of CSCs. Notes: Effect of the treatments on the proportion of CSCs in breast cancer cells, as reflected by the tumorsphere formation rate ( A and B ) and the proportion of ALDH+ cells ( E and F ). Representative images of tumorspheres formed by ( C ) MDA-MB-361 cells and ( D ) BT-474 cells are shown. The rate of tumorsphere formation is defined as the number of tumorspheres formed in 7 days in the treatment group divided by the number of tumorspheres formed in 7 days in the untreated group; the rate of tumorsphere formation in the untreated group is used as a control and defined as 100%. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=6). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSCs, cancer stem cells; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Journal: International Journal of Nanomedicine

    Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

    doi: 10.2147/IJN.S144184

    Figure Lengend Snippet: Effect of treatments on the percentage of CSCs. Notes: Effect of the treatments on the proportion of CSCs in breast cancer cells, as reflected by the tumorsphere formation rate ( A and B ) and the proportion of ALDH+ cells ( E and F ). Representative images of tumorspheres formed by ( C ) MDA-MB-361 cells and ( D ) BT-474 cells are shown. The rate of tumorsphere formation is defined as the number of tumorspheres formed in 7 days in the treatment group divided by the number of tumorspheres formed in 7 days in the untreated group; the rate of tumorsphere formation in the untreated group is used as a control and defined as 100%. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=6). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSCs, cancer stem cells; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Article Snippet: Human breast cancer cell lines BT-474 and MDA-MB-361 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques:

    In vivo antitumor assay in mice bearing subcutaneous BT-474 tumors. Notes: BT-474 tumors reached ~50 mm 3 in size by day 10. From day 10, mice were injected with nanoparticles (7.5 mg salinomycin/kg, i.v.) through the tail vein, and salinomycin (7.5 mg salinomycin/kg) dissolved in ethanol was administered by intraperitoneal injection. Therapy was given nine times on alternate days (indicated by arrows), and tumor volume was calculated. ( A ) Tumor growth curve. ( B ) Excised tumors. ( C ) The excised tumors were weighed at the end point. On day 28, the effect of the drug treatments on the CSC proportion in BT-474 tumors in vivo was evaluated by ( D ) the rate of tumorsphere formation and ( E ) the proportion of ALDH+ cells from the excised tumors. ( F ) Representative images of tumorspheres from ( D ) are shown. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=8). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSC, cancer stem cell; i.v., intravenous; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Journal: International Journal of Nanomedicine

    Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

    doi: 10.2147/IJN.S144184

    Figure Lengend Snippet: In vivo antitumor assay in mice bearing subcutaneous BT-474 tumors. Notes: BT-474 tumors reached ~50 mm 3 in size by day 10. From day 10, mice were injected with nanoparticles (7.5 mg salinomycin/kg, i.v.) through the tail vein, and salinomycin (7.5 mg salinomycin/kg) dissolved in ethanol was administered by intraperitoneal injection. Therapy was given nine times on alternate days (indicated by arrows), and tumor volume was calculated. ( A ) Tumor growth curve. ( B ) Excised tumors. ( C ) The excised tumors were weighed at the end point. On day 28, the effect of the drug treatments on the CSC proportion in BT-474 tumors in vivo was evaluated by ( D ) the rate of tumorsphere formation and ( E ) the proportion of ALDH+ cells from the excised tumors. ( F ) Representative images of tumorspheres from ( D ) are shown. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=8). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSC, cancer stem cell; i.v., intravenous; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

    Article Snippet: Human breast cancer cell lines BT-474 and MDA-MB-361 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: In Vivo, Injection

    Trastuzumab resistant cells were developed by culturing parental BT474 cells in the presence of 20/50 μ g trastuzumab for around 6 months.

    Journal: PLoS ONE

    Article Title: mRNA Profiling Reveals Determinants of Trastuzumab Efficiency in HER2-Positive Breast Cancer

    doi: 10.1371/journal.pone.0117818

    Figure Lengend Snippet: Trastuzumab resistant cells were developed by culturing parental BT474 cells in the presence of 20/50 μ g trastuzumab for around 6 months.

    Article Snippet: The human breast cancer cell line BT474 was directly obtained from the American Type Culture Collection (ATCC), catalogue no. HTB-20.

    Techniques:

    (a) Proliferation rate of BT474 (parental) and trastuzumab resistant BT474 (BTR50) cells treated with 20 μ g/ml trastuzumab. Proliferation rates were measured daily over 7 days by a luciferase-based viability assay. (b) Sensitivity of BT474 (parental) and trastuzumab resistant BTR50 cells towards increasing concentrations of trastuzumab. Cell viability was determined by a luciferase-based viability assay after 7 days of treatment.

    Journal: PLoS ONE

    Article Title: mRNA Profiling Reveals Determinants of Trastuzumab Efficiency in HER2-Positive Breast Cancer

    doi: 10.1371/journal.pone.0117818

    Figure Lengend Snippet: (a) Proliferation rate of BT474 (parental) and trastuzumab resistant BT474 (BTR50) cells treated with 20 μ g/ml trastuzumab. Proliferation rates were measured daily over 7 days by a luciferase-based viability assay. (b) Sensitivity of BT474 (parental) and trastuzumab resistant BTR50 cells towards increasing concentrations of trastuzumab. Cell viability was determined by a luciferase-based viability assay after 7 days of treatment.

    Article Snippet: The human breast cancer cell line BT474 was directly obtained from the American Type Culture Collection (ATCC), catalogue no. HTB-20.

    Techniques: Luciferase, Viability Assay

    The barchart displays the log2 fold changes of validated candidate genes, which significantly changed their expression in BT474 after trastuzumab treatment. The positive values indicate an upregulation upon drug treatment. Black bars denote values resulting from RNA-Seq analysis. Gray bars denote values resulting from RT-qPCR analysis.

    Journal: PLoS ONE

    Article Title: mRNA Profiling Reveals Determinants of Trastuzumab Efficiency in HER2-Positive Breast Cancer

    doi: 10.1371/journal.pone.0117818

    Figure Lengend Snippet: The barchart displays the log2 fold changes of validated candidate genes, which significantly changed their expression in BT474 after trastuzumab treatment. The positive values indicate an upregulation upon drug treatment. Black bars denote values resulting from RNA-Seq analysis. Gray bars denote values resulting from RT-qPCR analysis.

    Article Snippet: The human breast cancer cell line BT474 was directly obtained from the American Type Culture Collection (ATCC), catalogue no. HTB-20.

    Techniques: Expressing, RNA Sequencing Assay, Quantitative RT-PCR

    Upregulated genes in  BT474  upon trastuzumab treatment.

    Journal: PLoS ONE

    Article Title: mRNA Profiling Reveals Determinants of Trastuzumab Efficiency in HER2-Positive Breast Cancer

    doi: 10.1371/journal.pone.0117818

    Figure Lengend Snippet: Upregulated genes in BT474 upon trastuzumab treatment.

    Article Snippet: The human breast cancer cell line BT474 was directly obtained from the American Type Culture Collection (ATCC), catalogue no. HTB-20.

    Techniques: Binding Assay

    The barchart displays the log2 fold changes of validated candidate genes, which showed significant differences in their expression in BT474 and HCC1954, respectively. Positive values indicate an upregulation in BT474. Negative values indicate an upregulation in HCC1954. Black bars denote values resulting from RNA-Seq analysis. Gray bars denote values resulting from RT-qPCR analysis.

    Journal: PLoS ONE

    Article Title: mRNA Profiling Reveals Determinants of Trastuzumab Efficiency in HER2-Positive Breast Cancer

    doi: 10.1371/journal.pone.0117818

    Figure Lengend Snippet: The barchart displays the log2 fold changes of validated candidate genes, which showed significant differences in their expression in BT474 and HCC1954, respectively. Positive values indicate an upregulation in BT474. Negative values indicate an upregulation in HCC1954. Black bars denote values resulting from RNA-Seq analysis. Gray bars denote values resulting from RT-qPCR analysis.

    Article Snippet: The human breast cancer cell line BT474 was directly obtained from the American Type Culture Collection (ATCC), catalogue no. HTB-20.

    Techniques: Expressing, RNA Sequencing Assay, Quantitative RT-PCR

    Differentially expressed genes between HCC1954 and  BT474.

    Journal: PLoS ONE

    Article Title: mRNA Profiling Reveals Determinants of Trastuzumab Efficiency in HER2-Positive Breast Cancer

    doi: 10.1371/journal.pone.0117818

    Figure Lengend Snippet: Differentially expressed genes between HCC1954 and BT474.

    Article Snippet: The human breast cancer cell line BT474 was directly obtained from the American Type Culture Collection (ATCC), catalogue no. HTB-20.

    Techniques: Binding Assay

    The plot displays the result of a PCA on the pairwise normalized RNA-Seq expression values of all 23367 annotated genes in the samples BT474, BTR50 and HCC1954 with and without trastuzumab (T) treatment. Same colors denote that the samples belong to the same conducted statistical test and thus were normalized in pair. The five two sample tests included BT474 vs. HCC1954 , BT474 vs. BTR50 , HCC1954+T vs. HCC1954 , BTR50+T vs. BTR50 , and BT474+T vs. BT474 .

    Journal: PLoS ONE

    Article Title: mRNA Profiling Reveals Determinants of Trastuzumab Efficiency in HER2-Positive Breast Cancer

    doi: 10.1371/journal.pone.0117818

    Figure Lengend Snippet: The plot displays the result of a PCA on the pairwise normalized RNA-Seq expression values of all 23367 annotated genes in the samples BT474, BTR50 and HCC1954 with and without trastuzumab (T) treatment. Same colors denote that the samples belong to the same conducted statistical test and thus were normalized in pair. The five two sample tests included BT474 vs. HCC1954 , BT474 vs. BTR50 , HCC1954+T vs. HCC1954 , BTR50+T vs. BTR50 , and BT474+T vs. BT474 .

    Article Snippet: The human breast cancer cell line BT474 was directly obtained from the American Type Culture Collection (ATCC), catalogue no. HTB-20.

    Techniques: RNA Sequencing Assay, Expressing

    SNPs called in the  BT474  cell line.

    Journal: PLoS ONE

    Article Title: mRNA Profiling Reveals Determinants of Trastuzumab Efficiency in HER2-Positive Breast Cancer

    doi: 10.1371/journal.pone.0117818

    Figure Lengend Snippet: SNPs called in the BT474 cell line.

    Article Snippet: The human breast cancer cell line BT474 was directly obtained from the American Type Culture Collection (ATCC), catalogue no. HTB-20.

    Techniques:

    Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the BT474 breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)

    Journal: Molecular Cancer

    Article Title: Phosphorylation of Ser 78 of Hsp27 correlated with HER-2/ neu status and lymph node positivity in breast cancer

    doi: 10.1186/1476-4598-6-52

    Figure Lengend Snippet: Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the BT474 breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)

    Article Snippet: The human BT474 breast cancer cell line was obtained from American Type Culture Collection, and maintained in modified Dulbecco's medium (HybriCare) supplemented with 10% fetal bovine serum (FBS) at 37°C in a humidified atmosphere of 5% CO 2 in an incubator.

    Techniques: SDS Page, Cell Culture