human breast cancer cell lines bt 474  (ATCC)


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    ATCC human breast cancer cell lines bt 474
    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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    breast cancer bt 474 cells  (ATCC)


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    ATCC breast cancer bt 474 cells
    Breast Cancer Bt 474 Cells, 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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    breast cancer bt 474 cell lines  (ATCC)


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    ATCC breast cancer bt 474 cell lines
    Breast Cancer Bt 474 Cell Lines, 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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    breast cancer bt 474 cell lines  (ATCC)


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    ATCC breast cancer bt 474 cell lines
    In vitro binding specificity of ( A ) [ 99m Tc]Tc-Z AC12* -Z AC12* -GGGC and ( B ) [ 99m Tc]Tc-Z AC12* -Z Taq_3 -GGGC by SKOV-3 and <t>BT-474</t> cells. For the pre-saturation of B7-H3, a 200-fold molar excess of the same non-labelled Z AC12* -Z AC12* -GGGC and Z AC12* -Z Taq_3 -GGGC Affibody molecules was added to the dishes before adding the corresponding radiolabelled conjugate, respectively. Data are normalized to the average value of cell-associated radioactivity for non-blocked value for <t>BT474.</t> Asterisk (*) marks significant ( p < 0.05) difference. The data are presented as an average value from three samples ± SD
    Breast Cancer Bt 474 Cell Lines, 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 "Comparison of approaches for increasing affinity of affibody molecules for imaging of B7-H3: dimerization and affinity maturation"

    Article Title: Comparison of approaches for increasing affinity of affibody molecules for imaging of B7-H3: dimerization and affinity maturation

    Journal: EJNMMI Radiopharmacy and Chemistry

    doi: 10.1186/s41181-024-00261-3

    In vitro binding specificity of ( A ) [ 99m Tc]Tc-Z AC12* -Z AC12* -GGGC and ( B ) [ 99m Tc]Tc-Z AC12* -Z Taq_3 -GGGC by SKOV-3 and BT-474 cells. For the pre-saturation of B7-H3, a 200-fold molar excess of the same non-labelled Z AC12* -Z AC12* -GGGC and Z AC12* -Z Taq_3 -GGGC Affibody molecules was added to the dishes before adding the corresponding radiolabelled conjugate, respectively. Data are normalized to the average value of cell-associated radioactivity for non-blocked value for BT474. Asterisk (*) marks significant ( p < 0.05) difference. The data are presented as an average value from three samples ± SD
    Figure Legend Snippet: In vitro binding specificity of ( A ) [ 99m Tc]Tc-Z AC12* -Z AC12* -GGGC and ( B ) [ 99m Tc]Tc-Z AC12* -Z Taq_3 -GGGC by SKOV-3 and BT-474 cells. For the pre-saturation of B7-H3, a 200-fold molar excess of the same non-labelled Z AC12* -Z AC12* -GGGC and Z AC12* -Z Taq_3 -GGGC Affibody molecules was added to the dishes before adding the corresponding radiolabelled conjugate, respectively. Data are normalized to the average value of cell-associated radioactivity for non-blocked value for BT474. Asterisk (*) marks significant ( p < 0.05) difference. The data are presented as an average value from three samples ± SD

    Techniques Used: In Vitro, Binding Assay, Radioactivity

    breast cancer bt 474 cells  (ATCC)


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    ATCC breast cancer bt 474 cells
    Specificity of [ 177 Lu]Lu-labelled G3 binding in vitro to tumour cell lines without the addition of albumin. ( A ) [ 177 Lu]Lu-G3-ABD and ( B ) [ 177 Lu]Lu-ABD-G3 binding to SKOV-3 and <t>BT-474</t> (HER2-positive) and MDA-MB-468 (HER2-negative) cell lines. For the pre-saturation of HER2 receptors, an excess of a non-radioactive DARPin G3 was added before adding the labelled conjugate. ANOVA test (with Bonferroni’s correction for multiple comparisons) was performed to test if the difference was significant ( p < 0.05). Asterisk (*) marks significant ( p < 0.05) difference; n.s. that the difference is not significant.
    Breast Cancer Bt 474 Cells, 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 "Preclinical Evaluation of HER2-Targeting DARPin G3: Impact of Albumin-Binding Domain (ABD) Fusion"

    Article Title: Preclinical Evaluation of HER2-Targeting DARPin G3: Impact of Albumin-Binding Domain (ABD) Fusion

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms25084246

    Specificity of [ 177 Lu]Lu-labelled G3 binding in vitro to tumour cell lines without the addition of albumin. ( A ) [ 177 Lu]Lu-G3-ABD and ( B ) [ 177 Lu]Lu-ABD-G3 binding to SKOV-3 and BT-474 (HER2-positive) and MDA-MB-468 (HER2-negative) cell lines. For the pre-saturation of HER2 receptors, an excess of a non-radioactive DARPin G3 was added before adding the labelled conjugate. ANOVA test (with Bonferroni’s correction for multiple comparisons) was performed to test if the difference was significant ( p < 0.05). Asterisk (*) marks significant ( p < 0.05) difference; n.s. that the difference is not significant.
    Figure Legend Snippet: Specificity of [ 177 Lu]Lu-labelled G3 binding in vitro to tumour cell lines without the addition of albumin. ( A ) [ 177 Lu]Lu-G3-ABD and ( B ) [ 177 Lu]Lu-ABD-G3 binding to SKOV-3 and BT-474 (HER2-positive) and MDA-MB-468 (HER2-negative) cell lines. For the pre-saturation of HER2 receptors, an excess of a non-radioactive DARPin G3 was added before adding the labelled conjugate. ANOVA test (with Bonferroni’s correction for multiple comparisons) was performed to test if the difference was significant ( p < 0.05). Asterisk (*) marks significant ( p < 0.05) difference; n.s. that the difference is not significant.

    Techniques Used: Binding Assay, In Vitro

    Specificity of [ 177 Lu]Lu-labelled G3 binding in vitro to tumour cell lines in the presence of human serum albumin. ( A ) [ 177 Lu]Lu-G3-ABD and ( B ) [ 177 Lu]Lu -ABD-G3 binding in vitro to SKOV-3 and BT-474 (HER2-positive) and MDA-MB-468 (HER2-negative) cell lines in the presence of 100 nM human serum albumin. For the pre-saturation of HER2 receptors, an excess of a non-radioactive DARPin G3 was added before adding the labelled conjugate. ANOVA test (with Bonferroni’s correction for multiple comparisons) was performed to test if the difference was significant ( p < 0.05). Asterisk (*) marks a significant ( p < 0.05) difference.
    Figure Legend Snippet: Specificity of [ 177 Lu]Lu-labelled G3 binding in vitro to tumour cell lines in the presence of human serum albumin. ( A ) [ 177 Lu]Lu-G3-ABD and ( B ) [ 177 Lu]Lu -ABD-G3 binding in vitro to SKOV-3 and BT-474 (HER2-positive) and MDA-MB-468 (HER2-negative) cell lines in the presence of 100 nM human serum albumin. For the pre-saturation of HER2 receptors, an excess of a non-radioactive DARPin G3 was added before adding the labelled conjugate. ANOVA test (with Bonferroni’s correction for multiple comparisons) was performed to test if the difference was significant ( p < 0.05). Asterisk (*) marks a significant ( p < 0.05) difference.

    Techniques Used: Binding Assay, In Vitro

    Cellular processing of [ 177 Lu]Lu-labelled G3-ABD and ABD-G3. ( A , C , E ) Processing by SKOV-3 and ( B , D , F ) BT-474 cells. ( A , B ) show the binding and processing of [ 177 Lu]Lu-G3-ABD without the addition of albumin. ( C , D ) show the binding and processing of [ 177 Lu]Lu-G3-ABD with the addition of albumin. ( E , F ) show the binding and processing of [ 177 Lu]Lu-ABD-G3 with the addition of albumin. The cells were incubated with 2 nM of the conjugates.
    Figure Legend Snippet: Cellular processing of [ 177 Lu]Lu-labelled G3-ABD and ABD-G3. ( A , C , E ) Processing by SKOV-3 and ( B , D , F ) BT-474 cells. ( A , B ) show the binding and processing of [ 177 Lu]Lu-G3-ABD without the addition of albumin. ( C , D ) show the binding and processing of [ 177 Lu]Lu-G3-ABD with the addition of albumin. ( E , F ) show the binding and processing of [ 177 Lu]Lu-ABD-G3 with the addition of albumin. The cells were incubated with 2 nM of the conjugates.

    Techniques Used: Binding Assay, Incubation

    bt 474 breast cancer cells  (ATCC)


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    ATCC bt 474 breast cancer cells
    Bt 474 Breast Cancer Cells, 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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    breast cancer cell lines bt 474  (ATCC)


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    ATCC breast cancer cell lines bt 474
    PSPC1 inhibition synergizes with olaparib in BRCA1/2 -mutated PSPC1-expressing cells. ( A ) Correlation between PSPC1 expression and olaparib sensitivity, which was defined as IC 50 ≤ 3.2 µM, in BRCA1/2-mutated ovarian cancer cell lines using GDSC data. p -values were analyzed by Student’s t -test. ( B , C ) MTT assay showed that combined PSPC1 siRNA and olaparib enhanced the antiproliferative effects in ( B ) <t>BT−474</t> and ( C ) SNU−251 cells. Cells were treated with 5 nM of PSPC1 siRNA and indicated concentrations of olaparib and incubated for 72 h. ( D , E ) Apoptosis assay carried out by flow cytometry after staining with annexin V-APC/PI, representative scatter plots of PI ( y -axis) and annexin V ( x -axis). The number of total apoptotic cells were increased after the treatment with combined PSPC1 siRNA (5 nM) and olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) for 72 h in ( D ) BT−474 and ( E ) SNU−251 cells. The data shown here were representative of three independent experiments. The bar graphs depicted the average of total apoptotic cells of three independent experiments. p -values were calculated by one-way ANOVA analysis, indicating ** p < 0.01, *** p < 0.001, **** p < 0.0001. Data are presented as mean ± standard deviation from three independent experiments. ( F , G ) The expressions of apoptosis markers, caspase−3 and cleaved caspase−3 were analyzed with Western blot in ( F ) BT−474 and ( G ) SNU−251 cells. Cells were treated for 72 h with PSPC1 siRNA (5 nM), olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) and their combination. GAPDH was used as a loading control.
    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 "PSPC1 Inhibition Synergizes with Poly(ADP-ribose) Polymerase Inhibitors in a Preclinical Model of BRCA-Mutated Breast/Ovarian Cancer"

    Article Title: PSPC1 Inhibition Synergizes with Poly(ADP-ribose) Polymerase Inhibitors in a Preclinical Model of BRCA-Mutated Breast/Ovarian Cancer

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms242317086

    PSPC1 inhibition synergizes with olaparib in BRCA1/2 -mutated PSPC1-expressing cells. ( A ) Correlation between PSPC1 expression and olaparib sensitivity, which was defined as IC 50 ≤ 3.2 µM, in BRCA1/2-mutated ovarian cancer cell lines using GDSC data. p -values were analyzed by Student’s t -test. ( B , C ) MTT assay showed that combined PSPC1 siRNA and olaparib enhanced the antiproliferative effects in ( B ) BT−474 and ( C ) SNU−251 cells. Cells were treated with 5 nM of PSPC1 siRNA and indicated concentrations of olaparib and incubated for 72 h. ( D , E ) Apoptosis assay carried out by flow cytometry after staining with annexin V-APC/PI, representative scatter plots of PI ( y -axis) and annexin V ( x -axis). The number of total apoptotic cells were increased after the treatment with combined PSPC1 siRNA (5 nM) and olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) for 72 h in ( D ) BT−474 and ( E ) SNU−251 cells. The data shown here were representative of three independent experiments. The bar graphs depicted the average of total apoptotic cells of three independent experiments. p -values were calculated by one-way ANOVA analysis, indicating ** p < 0.01, *** p < 0.001, **** p < 0.0001. Data are presented as mean ± standard deviation from three independent experiments. ( F , G ) The expressions of apoptosis markers, caspase−3 and cleaved caspase−3 were analyzed with Western blot in ( F ) BT−474 and ( G ) SNU−251 cells. Cells were treated for 72 h with PSPC1 siRNA (5 nM), olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) and their combination. GAPDH was used as a loading control.
    Figure Legend Snippet: PSPC1 inhibition synergizes with olaparib in BRCA1/2 -mutated PSPC1-expressing cells. ( A ) Correlation between PSPC1 expression and olaparib sensitivity, which was defined as IC 50 ≤ 3.2 µM, in BRCA1/2-mutated ovarian cancer cell lines using GDSC data. p -values were analyzed by Student’s t -test. ( B , C ) MTT assay showed that combined PSPC1 siRNA and olaparib enhanced the antiproliferative effects in ( B ) BT−474 and ( C ) SNU−251 cells. Cells were treated with 5 nM of PSPC1 siRNA and indicated concentrations of olaparib and incubated for 72 h. ( D , E ) Apoptosis assay carried out by flow cytometry after staining with annexin V-APC/PI, representative scatter plots of PI ( y -axis) and annexin V ( x -axis). The number of total apoptotic cells were increased after the treatment with combined PSPC1 siRNA (5 nM) and olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) for 72 h in ( D ) BT−474 and ( E ) SNU−251 cells. The data shown here were representative of three independent experiments. The bar graphs depicted the average of total apoptotic cells of three independent experiments. p -values were calculated by one-way ANOVA analysis, indicating ** p < 0.01, *** p < 0.001, **** p < 0.0001. Data are presented as mean ± standard deviation from three independent experiments. ( F , G ) The expressions of apoptosis markers, caspase−3 and cleaved caspase−3 were analyzed with Western blot in ( F ) BT−474 and ( G ) SNU−251 cells. Cells were treated for 72 h with PSPC1 siRNA (5 nM), olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) and their combination. GAPDH was used as a loading control.

    Techniques Used: Inhibition, Expressing, MTT Assay, Incubation, Apoptosis Assay, Flow Cytometry, Staining, Concentration Assay, Standard Deviation, Western Blot

    PSPC1 inhibition enhances DNA DSBs by inhibiting olaparib-induced DDR. ( A , B ) Western blot was performed to analyze the expressions of DDR-related proteins in ( A ) BT−474 and ( B ) SNU−251 cells. Cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) and their combination for 72 h. GAPDH was used as a loading control. ( C , D ) Immunofluorescence was carried out to capture the ɣH2AX foci formation in ( C ) BT−474 and ( D ) SNU−251 cells as a DNA DSBs marker. The cells were incubated for 48 h after treatment with PSPC1 siRNA (5 nM), olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251), and their combination. The images were taken at 100x magnification. The images shown here were representative of three independent experiments. The bar graphs depicted the average foci number per cell of three independent experiments. The foci were counted in 10 cells for each group. P -values were calculated by one-way ANOVA analysis, indicating **** p < 0.0001. Data are presented as mean ± standard deviation from 3 independent experiments.
    Figure Legend Snippet: PSPC1 inhibition enhances DNA DSBs by inhibiting olaparib-induced DDR. ( A , B ) Western blot was performed to analyze the expressions of DDR-related proteins in ( A ) BT−474 and ( B ) SNU−251 cells. Cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) and their combination for 72 h. GAPDH was used as a loading control. ( C , D ) Immunofluorescence was carried out to capture the ɣH2AX foci formation in ( C ) BT−474 and ( D ) SNU−251 cells as a DNA DSBs marker. The cells were incubated for 48 h after treatment with PSPC1 siRNA (5 nM), olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251), and their combination. The images were taken at 100x magnification. The images shown here were representative of three independent experiments. The bar graphs depicted the average foci number per cell of three independent experiments. The foci were counted in 10 cells for each group. P -values were calculated by one-way ANOVA analysis, indicating **** p < 0.0001. Data are presented as mean ± standard deviation from 3 independent experiments.

    Techniques Used: Inhibition, Western Blot, Concentration Assay, Immunofluorescence, Marker, Incubation, Standard Deviation

    Mitotic catastrophe caused by combination treatment could explain the synergistic mechanisms. ( A , B ) BT−474 and SNU−251 cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251), and their combination for 72 h. Cell cycle distribution was assessed by flow cytometry after staining with PI. The histogram exhibited the distribution of cells in various phases (G0/G1, S, and G2/M) and the bar graphs represented the percentage of cells in G0/G1, S, and G2/M phases of cell cycle. ( C , D ) Western blot was carried out to check the expression of cell cycle progression-related proteins such as p-cdc25C and p-cdc2. ( C ) BT−474 and ( D ) SNU−251 cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT-474 and half IC 50 concentration in SNU−251), and their combination for 72 h. GAPDH was used as a loading control. ( E ) Graphical view of the proposed mechanism. Olaparib induced inhibitory phosphorylation of CDK1 causes mitotic exit which is reversed by PSPC1 inhibition (due to PSPC1 inhibition induced ATM and DNA-PKcs suppression), eventually premature mitotic entry occurred of the DNA-damaged cells, leading to mitotic catastrophe.
    Figure Legend Snippet: Mitotic catastrophe caused by combination treatment could explain the synergistic mechanisms. ( A , B ) BT−474 and SNU−251 cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251), and their combination for 72 h. Cell cycle distribution was assessed by flow cytometry after staining with PI. The histogram exhibited the distribution of cells in various phases (G0/G1, S, and G2/M) and the bar graphs represented the percentage of cells in G0/G1, S, and G2/M phases of cell cycle. ( C , D ) Western blot was carried out to check the expression of cell cycle progression-related proteins such as p-cdc25C and p-cdc2. ( C ) BT−474 and ( D ) SNU−251 cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT-474 and half IC 50 concentration in SNU−251), and their combination for 72 h. GAPDH was used as a loading control. ( E ) Graphical view of the proposed mechanism. Olaparib induced inhibitory phosphorylation of CDK1 causes mitotic exit which is reversed by PSPC1 inhibition (due to PSPC1 inhibition induced ATM and DNA-PKcs suppression), eventually premature mitotic entry occurred of the DNA-damaged cells, leading to mitotic catastrophe.

    Techniques Used: Concentration Assay, Flow Cytometry, Staining, Western Blot, Expressing, Inhibition

    Combination of PSPC1 siRNA and olaparib inhibits tumor growth in a PSPC1-expressing BRCA2-mutated breast cancer xenograft model. ( A ) Graphical view of drug treatment schedule and animal experimental procedure. ( B ) The body weights of mice exhibited that the treated doses were well tolerated as weight loss was not observed ( C ) Mean tumor growth inhibition curves in mice which were treated with olaparib, PSPC1 siRNA, and their combination. Tumor volumes were measured three times a week by a vernier caliper. Indicated p -values were calculated by one-way ANOVA analysis on day 16 of treatment, whereas ** p < 0.01, *** p < 0.001, **** p < 0.0001. ( D ) Individual tumor volumes were shown on day 16. Data are presented as the mean ± SEM. ( E ) Photo of the excised tumors of each group shown after sacrificing on day 16. The red dotted circles indicate complete regression of the tumor ( F ) Western blot using BT−474 xenografted tumors after 16 days of treatment. Combination treatment suppressed the expression of DDR-associated genes, such as p-ATM, DNA-PKcs, p-cdc25C, and p-cdc2 compared to monotherapy. ( G ) Western blot using BT−474 xenografted tumors after 16 days of treatment. The expression of the apoptosis marker cleaved caspase-3 was increased after combination treatment compared to monotherapy.
    Figure Legend Snippet: Combination of PSPC1 siRNA and olaparib inhibits tumor growth in a PSPC1-expressing BRCA2-mutated breast cancer xenograft model. ( A ) Graphical view of drug treatment schedule and animal experimental procedure. ( B ) The body weights of mice exhibited that the treated doses were well tolerated as weight loss was not observed ( C ) Mean tumor growth inhibition curves in mice which were treated with olaparib, PSPC1 siRNA, and their combination. Tumor volumes were measured three times a week by a vernier caliper. Indicated p -values were calculated by one-way ANOVA analysis on day 16 of treatment, whereas ** p < 0.01, *** p < 0.001, **** p < 0.0001. ( D ) Individual tumor volumes were shown on day 16. Data are presented as the mean ± SEM. ( E ) Photo of the excised tumors of each group shown after sacrificing on day 16. The red dotted circles indicate complete regression of the tumor ( F ) Western blot using BT−474 xenografted tumors after 16 days of treatment. Combination treatment suppressed the expression of DDR-associated genes, such as p-ATM, DNA-PKcs, p-cdc25C, and p-cdc2 compared to monotherapy. ( G ) Western blot using BT−474 xenografted tumors after 16 days of treatment. The expression of the apoptosis marker cleaved caspase-3 was increased after combination treatment compared to monotherapy.

    Techniques Used: Expressing, Inhibition, Western Blot, Marker

    breast cancer bt 474 cell lines  (ATCC)


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    ATCC breast cancer bt 474 cell lines
    Breast Cancer Bt 474 Cell Lines, 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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    human breast cancer cell line bt 474  (ATCC)


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    ATCC human breast cancer cell line bt 474
    Analysis of PPAB001 for simultaneous binding to CD47 and CD24 (A) Simultaneous binding of PPAB001 to recombinant CD47 and CD24 antigen by sandwich ELISA. (B) Flow cytometry of breast cancer <t>BT-474</t> cells stained with PPAB001, anti-CD24, CV1-hFc, or hIgG at 10 μg/mL. Binding kinetics of PPAB001, anti-CD24, or CV1-hFc to membrane-associated CD47 and CD24 on BT-474 cells were shown. Data are shown as means ± SDs (n = 3).
    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, 2024-07
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    1) Product Images from "Dual blockade of CD47 and CD24 signaling using a novel bispecific antibody fusion protein enhances macrophage immunotherapy"

    Article Title: Dual blockade of CD47 and CD24 signaling using a novel bispecific antibody fusion protein enhances macrophage immunotherapy

    Journal: Molecular Therapy Oncolytics

    doi: 10.1016/j.omto.2023.100747

    Analysis of PPAB001 for simultaneous binding to CD47 and CD24 (A) Simultaneous binding of PPAB001 to recombinant CD47 and CD24 antigen by sandwich ELISA. (B) Flow cytometry of breast cancer BT-474 cells stained with PPAB001, anti-CD24, CV1-hFc, or hIgG at 10 μg/mL. Binding kinetics of PPAB001, anti-CD24, or CV1-hFc to membrane-associated CD47 and CD24 on BT-474 cells were shown. Data are shown as means ± SDs (n = 3).
    Figure Legend Snippet: Analysis of PPAB001 for simultaneous binding to CD47 and CD24 (A) Simultaneous binding of PPAB001 to recombinant CD47 and CD24 antigen by sandwich ELISA. (B) Flow cytometry of breast cancer BT-474 cells stained with PPAB001, anti-CD24, CV1-hFc, or hIgG at 10 μg/mL. Binding kinetics of PPAB001, anti-CD24, or CV1-hFc to membrane-associated CD47 and CD24 on BT-474 cells were shown. Data are shown as means ± SDs (n = 3).

    Techniques Used: Binding Assay, Recombinant, Sandwich ELISA, Flow Cytometry, Staining

    Treatment with PPAB001 promotes phagocytosis of human cancer cells (A) Representative phagocytosis images of macrophages engulfing BT-474, BT-474R, or SK-OV-3 cells with PPAB001, anti-CD24, CV1-hFc, or hIgG. Macrophages were stained red (CMTPX); cancer cells were labeled with green (CFSE). The white arrows indicate macrophages that engulfed cancer cells. (B) The extent of macrophage phagocytosis with PPAB001, anti-CD24, CV1-hFc, or hIgG was quantified for BT-474, BT-474R, or SK-OV-3 cells. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.
    Figure Legend Snippet: Treatment with PPAB001 promotes phagocytosis of human cancer cells (A) Representative phagocytosis images of macrophages engulfing BT-474, BT-474R, or SK-OV-3 cells with PPAB001, anti-CD24, CV1-hFc, or hIgG. Macrophages were stained red (CMTPX); cancer cells were labeled with green (CFSE). The white arrows indicate macrophages that engulfed cancer cells. (B) The extent of macrophage phagocytosis with PPAB001, anti-CD24, CV1-hFc, or hIgG was quantified for BT-474, BT-474R, or SK-OV-3 cells. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

    Techniques Used: Staining, Labeling

    CD47/CD24 dual blockage potentiates the macrophage phagocytosis (A) Representative flow cytometry plots depicting the phagocytosis of BT-474 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (B) Representative flow cytometry plots depicting the phagocytosis of BT-474R cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (C) Representative flow cytometry plots depicting the phagocytosis of SK-OV-3 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.
    Figure Legend Snippet: CD47/CD24 dual blockage potentiates the macrophage phagocytosis (A) Representative flow cytometry plots depicting the phagocytosis of BT-474 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (B) Representative flow cytometry plots depicting the phagocytosis of BT-474R cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (C) Representative flow cytometry plots depicting the phagocytosis of SK-OV-3 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

    Techniques Used: Flow Cytometry

    human breast cancer bt 474 cell lines  (ATCC)


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    ATCC human breast cancer bt 474 cell lines
    Human Breast Cancer Bt 474 Cell Lines, 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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    ATCC human breast cancer cell lines bt 474
    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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    86
    ATCC breast cancer bt 474 cells
    Breast Cancer Bt 474 Cells, 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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    ATCC breast cancer bt 474 cell lines
    Breast Cancer Bt 474 Cell Lines, 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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    ATCC bt 474 breast cancer cells
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    ATCC breast cancer cell lines bt 474
    PSPC1 inhibition synergizes with olaparib in BRCA1/2 -mutated PSPC1-expressing cells. ( A ) Correlation between PSPC1 expression and olaparib sensitivity, which was defined as IC 50 ≤ 3.2 µM, in BRCA1/2-mutated ovarian cancer cell lines using GDSC data. p -values were analyzed by Student’s t -test. ( B , C ) MTT assay showed that combined PSPC1 siRNA and olaparib enhanced the antiproliferative effects in ( B ) <t>BT−474</t> and ( C ) SNU−251 cells. Cells were treated with 5 nM of PSPC1 siRNA and indicated concentrations of olaparib and incubated for 72 h. ( D , E ) Apoptosis assay carried out by flow cytometry after staining with annexin V-APC/PI, representative scatter plots of PI ( y -axis) and annexin V ( x -axis). The number of total apoptotic cells were increased after the treatment with combined PSPC1 siRNA (5 nM) and olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) for 72 h in ( D ) BT−474 and ( E ) SNU−251 cells. The data shown here were representative of three independent experiments. The bar graphs depicted the average of total apoptotic cells of three independent experiments. p -values were calculated by one-way ANOVA analysis, indicating ** p < 0.01, *** p < 0.001, **** p < 0.0001. Data are presented as mean ± standard deviation from three independent experiments. ( F , G ) The expressions of apoptosis markers, caspase−3 and cleaved caspase−3 were analyzed with Western blot in ( F ) BT−474 and ( G ) SNU−251 cells. Cells were treated for 72 h with PSPC1 siRNA (5 nM), olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) and their combination. GAPDH was used as a loading control.
    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/breast cancer cell lines bt 474/product/ATCC
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    86
    ATCC human breast cancer cell line bt 474
    Analysis of PPAB001 for simultaneous binding to CD47 and CD24 (A) Simultaneous binding of PPAB001 to recombinant CD47 and CD24 antigen by sandwich ELISA. (B) Flow cytometry of breast cancer <t>BT-474</t> cells stained with PPAB001, anti-CD24, CV1-hFc, or hIgG at 10 μg/mL. Binding kinetics of PPAB001, anti-CD24, or CV1-hFc to membrane-associated CD47 and CD24 on BT-474 cells were shown. Data are shown as means ± SDs (n = 3).
    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, 2024-07
    86/100 stars
      Buy from Supplier

    86
    ATCC human breast cancer bt 474 cell lines
    Analysis of PPAB001 for simultaneous binding to CD47 and CD24 (A) Simultaneous binding of PPAB001 to recombinant CD47 and CD24 antigen by sandwich ELISA. (B) Flow cytometry of breast cancer <t>BT-474</t> cells stained with PPAB001, anti-CD24, CV1-hFc, or hIgG at 10 μg/mL. Binding kinetics of PPAB001, anti-CD24, or CV1-hFc to membrane-associated CD47 and CD24 on BT-474 cells were shown. Data are shown as means ± SDs (n = 3).
    Human Breast Cancer Bt 474 Cell Lines, 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 bt 474 cell lines/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human breast cancer bt 474 cell lines - by Bioz Stars, 2024-07
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    PSPC1 inhibition synergizes with olaparib in BRCA1/2 -mutated PSPC1-expressing cells. ( A ) Correlation between PSPC1 expression and olaparib sensitivity, which was defined as IC 50 ≤ 3.2 µM, in BRCA1/2-mutated ovarian cancer cell lines using GDSC data. p -values were analyzed by Student’s t -test. ( B , C ) MTT assay showed that combined PSPC1 siRNA and olaparib enhanced the antiproliferative effects in ( B ) BT−474 and ( C ) SNU−251 cells. Cells were treated with 5 nM of PSPC1 siRNA and indicated concentrations of olaparib and incubated for 72 h. ( D , E ) Apoptosis assay carried out by flow cytometry after staining with annexin V-APC/PI, representative scatter plots of PI ( y -axis) and annexin V ( x -axis). The number of total apoptotic cells were increased after the treatment with combined PSPC1 siRNA (5 nM) and olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) for 72 h in ( D ) BT−474 and ( E ) SNU−251 cells. The data shown here were representative of three independent experiments. The bar graphs depicted the average of total apoptotic cells of three independent experiments. p -values were calculated by one-way ANOVA analysis, indicating ** p < 0.01, *** p < 0.001, **** p < 0.0001. Data are presented as mean ± standard deviation from three independent experiments. ( F , G ) The expressions of apoptosis markers, caspase−3 and cleaved caspase−3 were analyzed with Western blot in ( F ) BT−474 and ( G ) SNU−251 cells. Cells were treated for 72 h with PSPC1 siRNA (5 nM), olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) and their combination. GAPDH was used as a loading control.

    Journal: International Journal of Molecular Sciences

    Article Title: PSPC1 Inhibition Synergizes with Poly(ADP-ribose) Polymerase Inhibitors in a Preclinical Model of BRCA-Mutated Breast/Ovarian Cancer

    doi: 10.3390/ijms242317086

    Figure Lengend Snippet: PSPC1 inhibition synergizes with olaparib in BRCA1/2 -mutated PSPC1-expressing cells. ( A ) Correlation between PSPC1 expression and olaparib sensitivity, which was defined as IC 50 ≤ 3.2 µM, in BRCA1/2-mutated ovarian cancer cell lines using GDSC data. p -values were analyzed by Student’s t -test. ( B , C ) MTT assay showed that combined PSPC1 siRNA and olaparib enhanced the antiproliferative effects in ( B ) BT−474 and ( C ) SNU−251 cells. Cells were treated with 5 nM of PSPC1 siRNA and indicated concentrations of olaparib and incubated for 72 h. ( D , E ) Apoptosis assay carried out by flow cytometry after staining with annexin V-APC/PI, representative scatter plots of PI ( y -axis) and annexin V ( x -axis). The number of total apoptotic cells were increased after the treatment with combined PSPC1 siRNA (5 nM) and olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) for 72 h in ( D ) BT−474 and ( E ) SNU−251 cells. The data shown here were representative of three independent experiments. The bar graphs depicted the average of total apoptotic cells of three independent experiments. p -values were calculated by one-way ANOVA analysis, indicating ** p < 0.01, *** p < 0.001, **** p < 0.0001. Data are presented as mean ± standard deviation from three independent experiments. ( F , G ) The expressions of apoptosis markers, caspase−3 and cleaved caspase−3 were analyzed with Western blot in ( F ) BT−474 and ( G ) SNU−251 cells. Cells were treated for 72 h with PSPC1 siRNA (5 nM), olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) and their combination. GAPDH was used as a loading control.

    Article Snippet: The breast cancer cell lines BT−474 and MDA-MB−436 were bought from American Type Culture Collection (Manassas, VA, USA).

    Techniques: Inhibition, Expressing, MTT Assay, Incubation, Apoptosis Assay, Flow Cytometry, Staining, Concentration Assay, Standard Deviation, Western Blot

    PSPC1 inhibition enhances DNA DSBs by inhibiting olaparib-induced DDR. ( A , B ) Western blot was performed to analyze the expressions of DDR-related proteins in ( A ) BT−474 and ( B ) SNU−251 cells. Cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) and their combination for 72 h. GAPDH was used as a loading control. ( C , D ) Immunofluorescence was carried out to capture the ɣH2AX foci formation in ( C ) BT−474 and ( D ) SNU−251 cells as a DNA DSBs marker. The cells were incubated for 48 h after treatment with PSPC1 siRNA (5 nM), olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251), and their combination. The images were taken at 100x magnification. The images shown here were representative of three independent experiments. The bar graphs depicted the average foci number per cell of three independent experiments. The foci were counted in 10 cells for each group. P -values were calculated by one-way ANOVA analysis, indicating **** p < 0.0001. Data are presented as mean ± standard deviation from 3 independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: PSPC1 Inhibition Synergizes with Poly(ADP-ribose) Polymerase Inhibitors in a Preclinical Model of BRCA-Mutated Breast/Ovarian Cancer

    doi: 10.3390/ijms242317086

    Figure Lengend Snippet: PSPC1 inhibition enhances DNA DSBs by inhibiting olaparib-induced DDR. ( A , B ) Western blot was performed to analyze the expressions of DDR-related proteins in ( A ) BT−474 and ( B ) SNU−251 cells. Cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251) and their combination for 72 h. GAPDH was used as a loading control. ( C , D ) Immunofluorescence was carried out to capture the ɣH2AX foci formation in ( C ) BT−474 and ( D ) SNU−251 cells as a DNA DSBs marker. The cells were incubated for 48 h after treatment with PSPC1 siRNA (5 nM), olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251), and their combination. The images were taken at 100x magnification. The images shown here were representative of three independent experiments. The bar graphs depicted the average foci number per cell of three independent experiments. The foci were counted in 10 cells for each group. P -values were calculated by one-way ANOVA analysis, indicating **** p < 0.0001. Data are presented as mean ± standard deviation from 3 independent experiments.

    Article Snippet: The breast cancer cell lines BT−474 and MDA-MB−436 were bought from American Type Culture Collection (Manassas, VA, USA).

    Techniques: Inhibition, Western Blot, Concentration Assay, Immunofluorescence, Marker, Incubation, Standard Deviation

    Mitotic catastrophe caused by combination treatment could explain the synergistic mechanisms. ( A , B ) BT−474 and SNU−251 cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251), and their combination for 72 h. Cell cycle distribution was assessed by flow cytometry after staining with PI. The histogram exhibited the distribution of cells in various phases (G0/G1, S, and G2/M) and the bar graphs represented the percentage of cells in G0/G1, S, and G2/M phases of cell cycle. ( C , D ) Western blot was carried out to check the expression of cell cycle progression-related proteins such as p-cdc25C and p-cdc2. ( C ) BT−474 and ( D ) SNU−251 cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT-474 and half IC 50 concentration in SNU−251), and their combination for 72 h. GAPDH was used as a loading control. ( E ) Graphical view of the proposed mechanism. Olaparib induced inhibitory phosphorylation of CDK1 causes mitotic exit which is reversed by PSPC1 inhibition (due to PSPC1 inhibition induced ATM and DNA-PKcs suppression), eventually premature mitotic entry occurred of the DNA-damaged cells, leading to mitotic catastrophe.

    Journal: International Journal of Molecular Sciences

    Article Title: PSPC1 Inhibition Synergizes with Poly(ADP-ribose) Polymerase Inhibitors in a Preclinical Model of BRCA-Mutated Breast/Ovarian Cancer

    doi: 10.3390/ijms242317086

    Figure Lengend Snippet: Mitotic catastrophe caused by combination treatment could explain the synergistic mechanisms. ( A , B ) BT−474 and SNU−251 cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT−474 and half IC 50 concentration in SNU−251), and their combination for 72 h. Cell cycle distribution was assessed by flow cytometry after staining with PI. The histogram exhibited the distribution of cells in various phases (G0/G1, S, and G2/M) and the bar graphs represented the percentage of cells in G0/G1, S, and G2/M phases of cell cycle. ( C , D ) Western blot was carried out to check the expression of cell cycle progression-related proteins such as p-cdc25C and p-cdc2. ( C ) BT−474 and ( D ) SNU−251 cells were treated with 5 nM of PSPC1 siRNA, olaparib (IC 50 concentration in BT-474 and half IC 50 concentration in SNU−251), and their combination for 72 h. GAPDH was used as a loading control. ( E ) Graphical view of the proposed mechanism. Olaparib induced inhibitory phosphorylation of CDK1 causes mitotic exit which is reversed by PSPC1 inhibition (due to PSPC1 inhibition induced ATM and DNA-PKcs suppression), eventually premature mitotic entry occurred of the DNA-damaged cells, leading to mitotic catastrophe.

    Article Snippet: The breast cancer cell lines BT−474 and MDA-MB−436 were bought from American Type Culture Collection (Manassas, VA, USA).

    Techniques: Concentration Assay, Flow Cytometry, Staining, Western Blot, Expressing, Inhibition

    Combination of PSPC1 siRNA and olaparib inhibits tumor growth in a PSPC1-expressing BRCA2-mutated breast cancer xenograft model. ( A ) Graphical view of drug treatment schedule and animal experimental procedure. ( B ) The body weights of mice exhibited that the treated doses were well tolerated as weight loss was not observed ( C ) Mean tumor growth inhibition curves in mice which were treated with olaparib, PSPC1 siRNA, and their combination. Tumor volumes were measured three times a week by a vernier caliper. Indicated p -values were calculated by one-way ANOVA analysis on day 16 of treatment, whereas ** p < 0.01, *** p < 0.001, **** p < 0.0001. ( D ) Individual tumor volumes were shown on day 16. Data are presented as the mean ± SEM. ( E ) Photo of the excised tumors of each group shown after sacrificing on day 16. The red dotted circles indicate complete regression of the tumor ( F ) Western blot using BT−474 xenografted tumors after 16 days of treatment. Combination treatment suppressed the expression of DDR-associated genes, such as p-ATM, DNA-PKcs, p-cdc25C, and p-cdc2 compared to monotherapy. ( G ) Western blot using BT−474 xenografted tumors after 16 days of treatment. The expression of the apoptosis marker cleaved caspase-3 was increased after combination treatment compared to monotherapy.

    Journal: International Journal of Molecular Sciences

    Article Title: PSPC1 Inhibition Synergizes with Poly(ADP-ribose) Polymerase Inhibitors in a Preclinical Model of BRCA-Mutated Breast/Ovarian Cancer

    doi: 10.3390/ijms242317086

    Figure Lengend Snippet: Combination of PSPC1 siRNA and olaparib inhibits tumor growth in a PSPC1-expressing BRCA2-mutated breast cancer xenograft model. ( A ) Graphical view of drug treatment schedule and animal experimental procedure. ( B ) The body weights of mice exhibited that the treated doses were well tolerated as weight loss was not observed ( C ) Mean tumor growth inhibition curves in mice which were treated with olaparib, PSPC1 siRNA, and their combination. Tumor volumes were measured three times a week by a vernier caliper. Indicated p -values were calculated by one-way ANOVA analysis on day 16 of treatment, whereas ** p < 0.01, *** p < 0.001, **** p < 0.0001. ( D ) Individual tumor volumes were shown on day 16. Data are presented as the mean ± SEM. ( E ) Photo of the excised tumors of each group shown after sacrificing on day 16. The red dotted circles indicate complete regression of the tumor ( F ) Western blot using BT−474 xenografted tumors after 16 days of treatment. Combination treatment suppressed the expression of DDR-associated genes, such as p-ATM, DNA-PKcs, p-cdc25C, and p-cdc2 compared to monotherapy. ( G ) Western blot using BT−474 xenografted tumors after 16 days of treatment. The expression of the apoptosis marker cleaved caspase-3 was increased after combination treatment compared to monotherapy.

    Article Snippet: The breast cancer cell lines BT−474 and MDA-MB−436 were bought from American Type Culture Collection (Manassas, VA, USA).

    Techniques: Expressing, Inhibition, Western Blot, Marker

    Analysis of PPAB001 for simultaneous binding to CD47 and CD24 (A) Simultaneous binding of PPAB001 to recombinant CD47 and CD24 antigen by sandwich ELISA. (B) Flow cytometry of breast cancer BT-474 cells stained with PPAB001, anti-CD24, CV1-hFc, or hIgG at 10 μg/mL. Binding kinetics of PPAB001, anti-CD24, or CV1-hFc to membrane-associated CD47 and CD24 on BT-474 cells were shown. Data are shown as means ± SDs (n = 3).

    Journal: Molecular Therapy Oncolytics

    Article Title: Dual blockade of CD47 and CD24 signaling using a novel bispecific antibody fusion protein enhances macrophage immunotherapy

    doi: 10.1016/j.omto.2023.100747

    Figure Lengend Snippet: Analysis of PPAB001 for simultaneous binding to CD47 and CD24 (A) Simultaneous binding of PPAB001 to recombinant CD47 and CD24 antigen by sandwich ELISA. (B) Flow cytometry of breast cancer BT-474 cells stained with PPAB001, anti-CD24, CV1-hFc, or hIgG at 10 μg/mL. Binding kinetics of PPAB001, anti-CD24, or CV1-hFc to membrane-associated CD47 and CD24 on BT-474 cells were shown. Data are shown as means ± SDs (n = 3).

    Article Snippet: The human breast cancer cell line BT-474 and the ovarian cancer cell line SK-OV-3 were obtained from the American Type Culture Collection (ATCC).

    Techniques: Binding Assay, Recombinant, Sandwich ELISA, Flow Cytometry, Staining

    Treatment with PPAB001 promotes phagocytosis of human cancer cells (A) Representative phagocytosis images of macrophages engulfing BT-474, BT-474R, or SK-OV-3 cells with PPAB001, anti-CD24, CV1-hFc, or hIgG. Macrophages were stained red (CMTPX); cancer cells were labeled with green (CFSE). The white arrows indicate macrophages that engulfed cancer cells. (B) The extent of macrophage phagocytosis with PPAB001, anti-CD24, CV1-hFc, or hIgG was quantified for BT-474, BT-474R, or SK-OV-3 cells. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

    Journal: Molecular Therapy Oncolytics

    Article Title: Dual blockade of CD47 and CD24 signaling using a novel bispecific antibody fusion protein enhances macrophage immunotherapy

    doi: 10.1016/j.omto.2023.100747

    Figure Lengend Snippet: Treatment with PPAB001 promotes phagocytosis of human cancer cells (A) Representative phagocytosis images of macrophages engulfing BT-474, BT-474R, or SK-OV-3 cells with PPAB001, anti-CD24, CV1-hFc, or hIgG. Macrophages were stained red (CMTPX); cancer cells were labeled with green (CFSE). The white arrows indicate macrophages that engulfed cancer cells. (B) The extent of macrophage phagocytosis with PPAB001, anti-CD24, CV1-hFc, or hIgG was quantified for BT-474, BT-474R, or SK-OV-3 cells. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

    Article Snippet: The human breast cancer cell line BT-474 and the ovarian cancer cell line SK-OV-3 were obtained from the American Type Culture Collection (ATCC).

    Techniques: Staining, Labeling

    CD47/CD24 dual blockage potentiates the macrophage phagocytosis (A) Representative flow cytometry plots depicting the phagocytosis of BT-474 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (B) Representative flow cytometry plots depicting the phagocytosis of BT-474R cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (C) Representative flow cytometry plots depicting the phagocytosis of SK-OV-3 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

    Journal: Molecular Therapy Oncolytics

    Article Title: Dual blockade of CD47 and CD24 signaling using a novel bispecific antibody fusion protein enhances macrophage immunotherapy

    doi: 10.1016/j.omto.2023.100747

    Figure Lengend Snippet: CD47/CD24 dual blockage potentiates the macrophage phagocytosis (A) Representative flow cytometry plots depicting the phagocytosis of BT-474 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (B) Representative flow cytometry plots depicting the phagocytosis of BT-474R cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. (C) Representative flow cytometry plots depicting the phagocytosis of SK-OV-3 cells treated with PPAB001, anti-CD24 mAb, or CV1-hFc, compared with the IgG control. Phagocytosis index is shown as a bar graph. Data are shown as means ± SDs (n = 3), and statistical significance was determined by a Student’s t test. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

    Article Snippet: The human breast cancer cell line BT-474 and the ovarian cancer cell line SK-OV-3 were obtained from the American Type Culture Collection (ATCC).

    Techniques: Flow Cytometry