mcf 7 cells  (ATCC)


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    Structured Review

    ATCC mcf 7 cells
    Wnt/β-catenin signaling activation and Dkk1 expression in human breast cancer cells in culture. A. Wnt/β-catenin signaling in human breast cancer cells. Cytosolic free β-catenin from <t>MCF-7,</t> MDA-MB-231 and MDA-MB-231/bone cells
    Mcf 7 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Breast Cancer-derived Dickkopf1 Inhibits Osteoblast Differentiation and Osteoprotegerin Expression: Implication for Breast Cancer Osteolytic Bone Metastases"

    Article Title: Breast Cancer-derived Dickkopf1 Inhibits Osteoblast Differentiation and Osteoprotegerin Expression: Implication for Breast Cancer Osteolytic Bone Metastases

    Journal:

    doi: 10.1002/ijc.23625

    Wnt/β-catenin signaling activation and Dkk1 expression in human breast cancer cells in culture. A. Wnt/β-catenin signaling in human breast cancer cells. Cytosolic free β-catenin from MCF-7, MDA-MB-231 and MDA-MB-231/bone cells
    Figure Legend Snippet: Wnt/β-catenin signaling activation and Dkk1 expression in human breast cancer cells in culture. A. Wnt/β-catenin signaling in human breast cancer cells. Cytosolic free β-catenin from MCF-7, MDA-MB-231 and MDA-MB-231/bone cells

    Techniques Used: Activation Assay, Expressing, Multiple Displacement Amplification

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    ATCC mcf 7 cells
    Wnt/β-catenin signaling activation and Dkk1 expression in human breast cancer cells in culture. A. Wnt/β-catenin signaling in human breast cancer cells. Cytosolic free β-catenin from <t>MCF-7,</t> MDA-MB-231 and MDA-MB-231/bone cells
    Mcf 7 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mcf 7 cells/product/ATCC
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    90/100 stars
      Buy from Supplier

    90
    ATCC human breast mcf 7
    IL-24 activates protein kinase A (PKA) in a concentration-dependent manner. ( A ) <t>MCF-7</t> cells were treated for 72 h with Ad.IL-24 (25, 50, and 100 pfu per cell) or Ad.vector (100 pfu per cell). Cells were collected, protein purified, and subjected to Western blot analysis to detect phospho-PKA substrates, PKA-Cα subunits, and β-actin proteins. ( B ) MCF-7 cells were treated with Ad.vector (100 pfu/cell) or Ad.IL-24 (25, 50, and 100 pfu per cell) and then assayed for the production of cAMP after 20 h of treatment. Numbers represent mean cyclic adenosine monophosphate (cAMP) (nM) concentration after normalization to control. An average of three independent experiments is shown ± SE ( n = 9). ( C , D ) MCF-7 cells were treated for 72 h with Ad.vector (control) or Ad.IL-24 at 100 pfu/cell, and either untreated or treated with 10 μM H-89 for 72 h. Cells were collected, protein purified, and subjected to Western blot analysis to detect phospho-PKA substrates, PKA-Cα subunit, phospho-ATF4, total ATF4, and β-actin.
    Human Breast Mcf 7, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC mcf 7 breast cancer cell lines
    The effects of SDP on the cytokine secretion profile of <t>MCF-7</t> and MDA-MB-231. The SDP reduces the concentration of CSF1 in breast cancer cell lines culture supernatants. * Indicates a statistical significance compared with the control group ( P
    Mcf 7 Breast Cancer Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC drug treatment mcf 7
    ERK1/2 inhibition abrogates IR-induced G2/M checkpoint activation. <t>MCF-7</t> cells were treated with or without 1 μM DOX in the presence or absence of 50 μM U0126 for 2 hr and washed. The cells were incubated in regular growth medium for additional 22 hr, in the presence of 100 ng/ml nocodazole, and analyzed for mitotic cells by FACS, which contain both 4N -DNA content and Histone H3-Ser10 phosphorylation, as described in Materials and Methods . Upper panel: the histograms shown are representative FACS analyses for mitotic cells in samples treated with/without DOX in the presence or absence of U0126. The location of mitotic cells in each sample is indicated ( M ). Lower panel: the bar graph compares percentage of mitotic cells presented in the indicated cell samples. Results shown are representative of two separate experiments.
    Drug Treatment Mcf 7, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Wnt/β-catenin signaling activation and Dkk1 expression in human breast cancer cells in culture. A. Wnt/β-catenin signaling in human breast cancer cells. Cytosolic free β-catenin from MCF-7, MDA-MB-231 and MDA-MB-231/bone cells

    Journal:

    Article Title: Breast Cancer-derived Dickkopf1 Inhibits Osteoblast Differentiation and Osteoprotegerin Expression: Implication for Breast Cancer Osteolytic Bone Metastases

    doi: 10.1002/ijc.23625

    Figure Lengend Snippet: Wnt/β-catenin signaling activation and Dkk1 expression in human breast cancer cells in culture. A. Wnt/β-catenin signaling in human breast cancer cells. Cytosolic free β-catenin from MCF-7, MDA-MB-231 and MDA-MB-231/bone cells

    Article Snippet: MCF-7 cells, C2C12 cells, Wnt3A-secreting L cells, and control L cells were obtained from American Type Culture Collection.

    Techniques: Activation Assay, Expressing, Multiple Displacement Amplification

    IL-24 activates protein kinase A (PKA) in a concentration-dependent manner. ( A ) MCF-7 cells were treated for 72 h with Ad.IL-24 (25, 50, and 100 pfu per cell) or Ad.vector (100 pfu per cell). Cells were collected, protein purified, and subjected to Western blot analysis to detect phospho-PKA substrates, PKA-Cα subunits, and β-actin proteins. ( B ) MCF-7 cells were treated with Ad.vector (100 pfu/cell) or Ad.IL-24 (25, 50, and 100 pfu per cell) and then assayed for the production of cAMP after 20 h of treatment. Numbers represent mean cyclic adenosine monophosphate (cAMP) (nM) concentration after normalization to control. An average of three independent experiments is shown ± SE ( n = 9). ( C , D ) MCF-7 cells were treated for 72 h with Ad.vector (control) or Ad.IL-24 at 100 pfu/cell, and either untreated or treated with 10 μM H-89 for 72 h. Cells were collected, protein purified, and subjected to Western blot analysis to detect phospho-PKA substrates, PKA-Cα subunit, phospho-ATF4, total ATF4, and β-actin.

    Journal: International Journal of Molecular Sciences

    Article Title: IL-24 Promotes Apoptosis through cAMP-Dependent PKA Pathways in Human Breast Cancer Cells

    doi: 10.3390/ijms19113561

    Figure Lengend Snippet: IL-24 activates protein kinase A (PKA) in a concentration-dependent manner. ( A ) MCF-7 cells were treated for 72 h with Ad.IL-24 (25, 50, and 100 pfu per cell) or Ad.vector (100 pfu per cell). Cells were collected, protein purified, and subjected to Western blot analysis to detect phospho-PKA substrates, PKA-Cα subunits, and β-actin proteins. ( B ) MCF-7 cells were treated with Ad.vector (100 pfu/cell) or Ad.IL-24 (25, 50, and 100 pfu per cell) and then assayed for the production of cAMP after 20 h of treatment. Numbers represent mean cyclic adenosine monophosphate (cAMP) (nM) concentration after normalization to control. An average of three independent experiments is shown ± SE ( n = 9). ( C , D ) MCF-7 cells were treated for 72 h with Ad.vector (control) or Ad.IL-24 at 100 pfu/cell, and either untreated or treated with 10 μM H-89 for 72 h. Cells were collected, protein purified, and subjected to Western blot analysis to detect phospho-PKA substrates, PKA-Cα subunit, phospho-ATF4, total ATF4, and β-actin.

    Article Snippet: Human breast MCF-7, MDA-MB-231, MDA-MB-157, and T47D cell lines were acquired from American Type Culture Collection (ATCC), maintained per ATCC protocols and utilized within six months of thawing.

    Techniques: Concentration Assay, Plasmid Preparation, Purification, Western Blot

    IL-24 induces TP53 expression, and promotes nuclear translocation in a PKA-dependent manner. ( A , B ) MCF-7 cells were treated for 72 h with Ad.IL-24 (25, 50, and 100 pfu per cell), 10 μM H-89, or Ad.vector (100 pfu per cell). Cells were collected, protein purified, and subjected to Western blot analysis to detect to detect phospho-TP53, total TP53, and β-actin proteins. ( C ) Semi-quantitative measurements of the mean fluorescein isothiocyanate (FITC) intensity were taken using Nikon NIS Elements whereby, the average intensity of five cell nuclei were taken under each experimental condition. Error bars are expressed as the standard deviation of FITC intensity values. ( D ) Cells were fixed and phospho-Ser15 TP53 was detected by immunofluorescence using anti-phospho-TP53 antibodies.

    Journal: International Journal of Molecular Sciences

    Article Title: IL-24 Promotes Apoptosis through cAMP-Dependent PKA Pathways in Human Breast Cancer Cells

    doi: 10.3390/ijms19113561

    Figure Lengend Snippet: IL-24 induces TP53 expression, and promotes nuclear translocation in a PKA-dependent manner. ( A , B ) MCF-7 cells were treated for 72 h with Ad.IL-24 (25, 50, and 100 pfu per cell), 10 μM H-89, or Ad.vector (100 pfu per cell). Cells were collected, protein purified, and subjected to Western blot analysis to detect to detect phospho-TP53, total TP53, and β-actin proteins. ( C ) Semi-quantitative measurements of the mean fluorescein isothiocyanate (FITC) intensity were taken using Nikon NIS Elements whereby, the average intensity of five cell nuclei were taken under each experimental condition. Error bars are expressed as the standard deviation of FITC intensity values. ( D ) Cells were fixed and phospho-Ser15 TP53 was detected by immunofluorescence using anti-phospho-TP53 antibodies.

    Article Snippet: Human breast MCF-7, MDA-MB-231, MDA-MB-157, and T47D cell lines were acquired from American Type Culture Collection (ATCC), maintained per ATCC protocols and utilized within six months of thawing.

    Techniques: Expressing, Translocation Assay, Plasmid Preparation, Purification, Western Blot, Standard Deviation, Immunofluorescence

    The inhibition of PKA blocks IL-24 activation of extrinsic apoptosis. ( A ) MCF-7 cells were infected with either Ad.vector (control) or increasing concentrations of Ad.IL-24 (25, 50, 100 pfu per cell) for 72 h. Western blot analysis was performed with antibodies for FasL, Fas, FADD, DR4, and β-actin. ( B ) MCF-7 cells were infected with either the Ad.vector (control) or Ad.IL-24 at 100 pfu/cell, and either untreated or treated with 10 μM H-89 for 72 h. Western blot analysis was performed with antibodies for FasL, Fas, FADD, DR4, and β-actin.

    Journal: International Journal of Molecular Sciences

    Article Title: IL-24 Promotes Apoptosis through cAMP-Dependent PKA Pathways in Human Breast Cancer Cells

    doi: 10.3390/ijms19113561

    Figure Lengend Snippet: The inhibition of PKA blocks IL-24 activation of extrinsic apoptosis. ( A ) MCF-7 cells were infected with either Ad.vector (control) or increasing concentrations of Ad.IL-24 (25, 50, 100 pfu per cell) for 72 h. Western blot analysis was performed with antibodies for FasL, Fas, FADD, DR4, and β-actin. ( B ) MCF-7 cells were infected with either the Ad.vector (control) or Ad.IL-24 at 100 pfu/cell, and either untreated or treated with 10 μM H-89 for 72 h. Western blot analysis was performed with antibodies for FasL, Fas, FADD, DR4, and β-actin.

    Article Snippet: Human breast MCF-7, MDA-MB-231, MDA-MB-157, and T47D cell lines were acquired from American Type Culture Collection (ATCC), maintained per ATCC protocols and utilized within six months of thawing.

    Techniques: Inhibition, Activation Assay, Infection, Plasmid Preparation, Western Blot

    The IL-24 killing effect is decreased in the presence of PKA inhibitors. ( A ) Human breast MCF-7, MDA-MB-231, MDA-MB-157, and T47D cancer cells were incubated with 10 μM of PKA inhibitor, H-89, or PKI, with or without Ad.IL-24 (100 pfu per cell) or Ad.vector (100 pfu per cell), and cell viability was determined by the MTT proliferation assay five days after treatment. Numbers represent the ratio of specific treatments to values in control cells (Ad.vector). An average of three independent experiments is shown ± SD as errors bars. *, p

    Journal: International Journal of Molecular Sciences

    Article Title: IL-24 Promotes Apoptosis through cAMP-Dependent PKA Pathways in Human Breast Cancer Cells

    doi: 10.3390/ijms19113561

    Figure Lengend Snippet: The IL-24 killing effect is decreased in the presence of PKA inhibitors. ( A ) Human breast MCF-7, MDA-MB-231, MDA-MB-157, and T47D cancer cells were incubated with 10 μM of PKA inhibitor, H-89, or PKI, with or without Ad.IL-24 (100 pfu per cell) or Ad.vector (100 pfu per cell), and cell viability was determined by the MTT proliferation assay five days after treatment. Numbers represent the ratio of specific treatments to values in control cells (Ad.vector). An average of three independent experiments is shown ± SD as errors bars. *, p

    Article Snippet: Human breast MCF-7, MDA-MB-231, MDA-MB-157, and T47D cell lines were acquired from American Type Culture Collection (ATCC), maintained per ATCC protocols and utilized within six months of thawing.

    Techniques: Multiple Displacement Amplification, Incubation, Plasmid Preparation, MTT Assay, Proliferation Assay

    IL-24 activates ATF4 in a dosage dependent manner. MCF-7 cells were treated for 72 h with Ad.IL-24 (25, 50, and 100 plaque-forming units (pfu) per cell) or Ad.vector (100 pfu per cell). Cells were collected, protein purified, and subjected to Western blot analysis to detect phospho-ATF4, total ATF4, BiP, and β-actin.

    Journal: International Journal of Molecular Sciences

    Article Title: IL-24 Promotes Apoptosis through cAMP-Dependent PKA Pathways in Human Breast Cancer Cells

    doi: 10.3390/ijms19113561

    Figure Lengend Snippet: IL-24 activates ATF4 in a dosage dependent manner. MCF-7 cells were treated for 72 h with Ad.IL-24 (25, 50, and 100 plaque-forming units (pfu) per cell) or Ad.vector (100 pfu per cell). Cells were collected, protein purified, and subjected to Western blot analysis to detect phospho-ATF4, total ATF4, BiP, and β-actin.

    Article Snippet: Human breast MCF-7, MDA-MB-231, MDA-MB-157, and T47D cell lines were acquired from American Type Culture Collection (ATCC), maintained per ATCC protocols and utilized within six months of thawing.

    Techniques: Plasmid Preparation, Purification, Western Blot

    The effects of SDP on the cytokine secretion profile of MCF-7 and MDA-MB-231. The SDP reduces the concentration of CSF1 in breast cancer cell lines culture supernatants. * Indicates a statistical significance compared with the control group ( P

    Journal: OncoTargets and therapy

    Article Title: Polysaccharides Derived from Saposhnikovia divaricata May Suppress Breast Cancer Through Activating Macrophages

    doi: 10.2147/OTT.S267984

    Figure Lengend Snippet: The effects of SDP on the cytokine secretion profile of MCF-7 and MDA-MB-231. The SDP reduces the concentration of CSF1 in breast cancer cell lines culture supernatants. * Indicates a statistical significance compared with the control group ( P

    Article Snippet: Cell Lines and ReagentsMDA-MB-231 and MCF-7 breast cancer cell lines were maintained in our institute, which were originally obtained from the American Type Culture Collection (ATCC).

    Techniques: Multiple Displacement Amplification, Concentration Assay

    The effects of SDP on immunosuppression by breast cancer cells. ( A ) The SDP antagonized the immunosuppression by breast cancer cells on U937 after co-culture with MCF-7 or MDA-MB-231 breast cancer cell cultural supernatants (CS). ( B ) The CD163 mRNA expression level was increased in U937 cells after co-culture with MCF-7 or MDA-MB-231 breast cancer cell CS, while SDP attenuated the effect. * Indicates a statistical significance compared with the control group ( P

    Journal: OncoTargets and therapy

    Article Title: Polysaccharides Derived from Saposhnikovia divaricata May Suppress Breast Cancer Through Activating Macrophages

    doi: 10.2147/OTT.S267984

    Figure Lengend Snippet: The effects of SDP on immunosuppression by breast cancer cells. ( A ) The SDP antagonized the immunosuppression by breast cancer cells on U937 after co-culture with MCF-7 or MDA-MB-231 breast cancer cell cultural supernatants (CS). ( B ) The CD163 mRNA expression level was increased in U937 cells after co-culture with MCF-7 or MDA-MB-231 breast cancer cell CS, while SDP attenuated the effect. * Indicates a statistical significance compared with the control group ( P

    Article Snippet: Cell Lines and ReagentsMDA-MB-231 and MCF-7 breast cancer cell lines were maintained in our institute, which were originally obtained from the American Type Culture Collection (ATCC).

    Techniques: Co-Culture Assay, Multiple Displacement Amplification, Expressing

    The proliferation effects of polysaccharides isolated from Saposhnikovia divaricata on U937 ( A ), MCF-7 ( B ), and MDA-MB-231 ( C ). SDP significantly stimulated the proliferation of U937 cells in a concentration dependent manner, while concentrations of these polysaccharides up to 800 μg/mL did not exhibit obvious cytotoxic effects on breast cancer cell lines in vitro. Error bars indicate standard deviation of the means. Data are exhibited as mean±SE of three independent results. * Indicates a statistical significance compared with the control group ( P

    Journal: OncoTargets and therapy

    Article Title: Polysaccharides Derived from Saposhnikovia divaricata May Suppress Breast Cancer Through Activating Macrophages

    doi: 10.2147/OTT.S267984

    Figure Lengend Snippet: The proliferation effects of polysaccharides isolated from Saposhnikovia divaricata on U937 ( A ), MCF-7 ( B ), and MDA-MB-231 ( C ). SDP significantly stimulated the proliferation of U937 cells in a concentration dependent manner, while concentrations of these polysaccharides up to 800 μg/mL did not exhibit obvious cytotoxic effects on breast cancer cell lines in vitro. Error bars indicate standard deviation of the means. Data are exhibited as mean±SE of three independent results. * Indicates a statistical significance compared with the control group ( P

    Article Snippet: Cell Lines and ReagentsMDA-MB-231 and MCF-7 breast cancer cell lines were maintained in our institute, which were originally obtained from the American Type Culture Collection (ATCC).

    Techniques: Isolation, Multiple Displacement Amplification, Concentration Assay, In Vitro, Standard Deviation

    The SDP antagonized TAMs recruitment by breast cancer cells in human breast cancer xenografts mice. The prevalence of CD163 positive macrophage among SDP treated and control mice through immunohistochemical analysis. As the figures indicates, the SDP antagonized TAMs recruitment by breast cancer cells in human breast cancer xenografts mice. ( A ) MDA-MB-231 blank control; ( B ) 200 μg/g SDP; ( C ) 400 μg/g SDP; ( D ) 800 μg/g SDP; and ( E ) MCF-7 control.

    Journal: OncoTargets and therapy

    Article Title: Polysaccharides Derived from Saposhnikovia divaricata May Suppress Breast Cancer Through Activating Macrophages

    doi: 10.2147/OTT.S267984

    Figure Lengend Snippet: The SDP antagonized TAMs recruitment by breast cancer cells in human breast cancer xenografts mice. The prevalence of CD163 positive macrophage among SDP treated and control mice through immunohistochemical analysis. As the figures indicates, the SDP antagonized TAMs recruitment by breast cancer cells in human breast cancer xenografts mice. ( A ) MDA-MB-231 blank control; ( B ) 200 μg/g SDP; ( C ) 400 μg/g SDP; ( D ) 800 μg/g SDP; and ( E ) MCF-7 control.

    Article Snippet: Cell Lines and ReagentsMDA-MB-231 and MCF-7 breast cancer cell lines were maintained in our institute, which were originally obtained from the American Type Culture Collection (ATCC).

    Techniques: Mouse Assay, Immunohistochemistry, Multiple Displacement Amplification

    The effects of SDP on tumor growth in MDA-MB-231 breast cancer xenografts. MDA-MB-231 tumor-bearing mice were randomly divided into an untreated group as the control and three treatment groups (200, 400, and 800 μg/g SDP), and the weak tumorigenic breast cancer cell line MCF-7 was also used as a control, with five mice per group. Tumor diameters and body weights of the mice in each group were measured with digital calipers and recorded each week. The tumor volume was computed as (width 2 ×length)/2 the tumor-bearing mice. The SDP suppressed tumor growth in both dose-dependent and time-dependent manners, the tumor volume ( A ) and tumor growth curves ( B ). * Indicates a statistical significance compared with the control group ( P

    Journal: OncoTargets and therapy

    Article Title: Polysaccharides Derived from Saposhnikovia divaricata May Suppress Breast Cancer Through Activating Macrophages

    doi: 10.2147/OTT.S267984

    Figure Lengend Snippet: The effects of SDP on tumor growth in MDA-MB-231 breast cancer xenografts. MDA-MB-231 tumor-bearing mice were randomly divided into an untreated group as the control and three treatment groups (200, 400, and 800 μg/g SDP), and the weak tumorigenic breast cancer cell line MCF-7 was also used as a control, with five mice per group. Tumor diameters and body weights of the mice in each group were measured with digital calipers and recorded each week. The tumor volume was computed as (width 2 ×length)/2 the tumor-bearing mice. The SDP suppressed tumor growth in both dose-dependent and time-dependent manners, the tumor volume ( A ) and tumor growth curves ( B ). * Indicates a statistical significance compared with the control group ( P

    Article Snippet: Cell Lines and ReagentsMDA-MB-231 and MCF-7 breast cancer cell lines were maintained in our institute, which were originally obtained from the American Type Culture Collection (ATCC).

    Techniques: Multiple Displacement Amplification, Mouse Assay

    ERK1/2 inhibition abrogates IR-induced G2/M checkpoint activation. MCF-7 cells were treated with or without 1 μM DOX in the presence or absence of 50 μM U0126 for 2 hr and washed. The cells were incubated in regular growth medium for additional 22 hr, in the presence of 100 ng/ml nocodazole, and analyzed for mitotic cells by FACS, which contain both 4N -DNA content and Histone H3-Ser10 phosphorylation, as described in Materials and Methods . Upper panel: the histograms shown are representative FACS analyses for mitotic cells in samples treated with/without DOX in the presence or absence of U0126. The location of mitotic cells in each sample is indicated ( M ). Lower panel: the bar graph compares percentage of mitotic cells presented in the indicated cell samples. Results shown are representative of two separate experiments.

    Journal: PLoS ONE

    Article Title: ERK1/2 Signaling Plays an Important Role in Topoisomerase II Poison-Induced G2/M Checkpoint Activation

    doi: 10.1371/journal.pone.0050281

    Figure Lengend Snippet: ERK1/2 inhibition abrogates IR-induced G2/M checkpoint activation. MCF-7 cells were treated with or without 1 μM DOX in the presence or absence of 50 μM U0126 for 2 hr and washed. The cells were incubated in regular growth medium for additional 22 hr, in the presence of 100 ng/ml nocodazole, and analyzed for mitotic cells by FACS, which contain both 4N -DNA content and Histone H3-Ser10 phosphorylation, as described in Materials and Methods . Upper panel: the histograms shown are representative FACS analyses for mitotic cells in samples treated with/without DOX in the presence or absence of U0126. The location of mitotic cells in each sample is indicated ( M ). Lower panel: the bar graph compares percentage of mitotic cells presented in the indicated cell samples. Results shown are representative of two separate experiments.

    Article Snippet: Cell culture and drug treatment MCF-7 and T47D human breast cancer cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS).

    Techniques: Inhibition, Activation Assay, Incubation, FACS

    Decrease of ATR level by shRNA had no effect on DOX-induced ERK1/2 activation. MCF-7 cells expressing ATR specific or control shRNA were treated with or without 1 µM DOX (upper panel) or 10 µM ETOP (lower panel) for 2 hr and analyzed for phospho-ERK1/2 ( pERK1/2 ) and total ERK1/2 ( ERK1/2 ) by immunoblotting.

    Journal: PLoS ONE

    Article Title: ERK1/2 Signaling Plays an Important Role in Topoisomerase II Poison-Induced G2/M Checkpoint Activation

    doi: 10.1371/journal.pone.0050281

    Figure Lengend Snippet: Decrease of ATR level by shRNA had no effect on DOX-induced ERK1/2 activation. MCF-7 cells expressing ATR specific or control shRNA were treated with or without 1 µM DOX (upper panel) or 10 µM ETOP (lower panel) for 2 hr and analyzed for phospho-ERK1/2 ( pERK1/2 ) and total ERK1/2 ( ERK1/2 ) by immunoblotting.

    Article Snippet: Cell culture and drug treatment MCF-7 and T47D human breast cancer cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS).

    Techniques: shRNA, Activation Assay, Expressing

    Inhibition of ERK1/2 signaling increases topo II poison-induced apoptosis. (A) MCF-7 cells were treated for 2 hr with or without 1 µM DOX or 10 µM ETOP in the presence or absence of 50 µM U0126. Following treatment, the cells were washed, incubated for 3 days with/without presence of U0126 and analyzed for apoptosis by DAPI staining and fluorescence microcopy. Upper panels: images shown are the DAPI staining of the resulting cells. Lower panels: the percentage of apoptotic cells is shown as mean ± s.d of quadruplicate samples. * p

    Journal: PLoS ONE

    Article Title: ERK1/2 Signaling Plays an Important Role in Topoisomerase II Poison-Induced G2/M Checkpoint Activation

    doi: 10.1371/journal.pone.0050281

    Figure Lengend Snippet: Inhibition of ERK1/2 signaling increases topo II poison-induced apoptosis. (A) MCF-7 cells were treated for 2 hr with or without 1 µM DOX or 10 µM ETOP in the presence or absence of 50 µM U0126. Following treatment, the cells were washed, incubated for 3 days with/without presence of U0126 and analyzed for apoptosis by DAPI staining and fluorescence microcopy. Upper panels: images shown are the DAPI staining of the resulting cells. Lower panels: the percentage of apoptotic cells is shown as mean ± s.d of quadruplicate samples. * p

    Article Snippet: Cell culture and drug treatment MCF-7 and T47D human breast cancer cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS).

    Techniques: Inhibition, Incubation, Staining, Fluorescence

    DOX and ETOP induce activation of Chk1 and Chk2 kinases and inhibition of Cdc2 kinase. (A) MCF-7 Cells were incubated with 1 µM DOX (upper panel) or 10 µM ETOP (lower panel) for the indicated times for up to 2 hr (lanes 1–4). For the 4 hr and 6 hr time points (lanes 5–6), the cells were incubated for 2 hr with DOX or ETOP, washed with DMEM and incubated for additional 2 hr and 4 hr, respectively, in regular culture medium. Following treatment, Chk1 and Chk2 kinases were respectively immunoprecipitated from cell lysates and examined for kinase activity as described in Materials and Methods ( Chk1 Activity and Chk2 Activity ). Levels of Chk1 and Chk2 in the immunoprecipitates were determined by immunoblotting ( Chk1 IP-WB and Chk2 IP-WB ). *, as a negative control, kinase assay was carried out using immunoprecipitates obtained by incubating DOX-treated cell sample (6 hr time point) with non-immunized IgG. (B) Cells were treated as described above and incubated for 2 hr. Cdc2 was immunoprecipitated from cell lysate and analyzed for levels of Cdc2-Tyr15 phosphorylation by immunoblotting ( Cdc2-Tyr15 ). As a control, Cdc2 in the immunoprecipitates was assessed by immunoblotting ( Cdc2 ).

    Journal: PLoS ONE

    Article Title: ERK1/2 Signaling Plays an Important Role in Topoisomerase II Poison-Induced G2/M Checkpoint Activation

    doi: 10.1371/journal.pone.0050281

    Figure Lengend Snippet: DOX and ETOP induce activation of Chk1 and Chk2 kinases and inhibition of Cdc2 kinase. (A) MCF-7 Cells were incubated with 1 µM DOX (upper panel) or 10 µM ETOP (lower panel) for the indicated times for up to 2 hr (lanes 1–4). For the 4 hr and 6 hr time points (lanes 5–6), the cells were incubated for 2 hr with DOX or ETOP, washed with DMEM and incubated for additional 2 hr and 4 hr, respectively, in regular culture medium. Following treatment, Chk1 and Chk2 kinases were respectively immunoprecipitated from cell lysates and examined for kinase activity as described in Materials and Methods ( Chk1 Activity and Chk2 Activity ). Levels of Chk1 and Chk2 in the immunoprecipitates were determined by immunoblotting ( Chk1 IP-WB and Chk2 IP-WB ). *, as a negative control, kinase assay was carried out using immunoprecipitates obtained by incubating DOX-treated cell sample (6 hr time point) with non-immunized IgG. (B) Cells were treated as described above and incubated for 2 hr. Cdc2 was immunoprecipitated from cell lysate and analyzed for levels of Cdc2-Tyr15 phosphorylation by immunoblotting ( Cdc2-Tyr15 ). As a control, Cdc2 in the immunoprecipitates was assessed by immunoblotting ( Cdc2 ).

    Article Snippet: Cell culture and drug treatment MCF-7 and T47D human breast cancer cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS).

    Techniques: Activation Assay, Inhibition, Incubation, Immunoprecipitation, Activity Assay, Western Blot, Negative Control, Kinase Assay

    DOX-induced G2/M checkpoint activation does not involve the p38 and MK2 kinases. (A) MCF-7 cells were treated for 2 hr with or without 1 μM DOX in the presence or absence of 50 μM U0126. As a positive control for p38 activation, a cell sample was exposed to UV at 100 J/m 2 and incubated for 1 hr at 37°C. The resulting cells were analyzed for levels of phospho-p38 ( p-p38 ) and total p38 ( p38 ) by immunoblotting. (B) The cell samples obtained above were analyzed for levels of phospho-MK2 ( pMK2 ), total MK2 ( MK2 ), phospho-ERK1/2 ( pERK1/2 ) and total ERK1/2 ( ERK1/2 ) by immunoblotting. (C) MCF-7 cells were transfected with control ( Control ) or MK2 specific siRNA ( MK2 ) and incubated for 2 days at 37°C. Upper panel: levels of MK2 and Actin in the transfected cells were determined by immunoblotting. Lower panel: The transfected cells were treated with 1 μM DOX, incubated for 24 hr and analyzed for DNA content by FACS. Graph depicts percentage of cells with 4N -DNA content and represents the mean ± s.d of two separate experiments with duplicate samples.

    Journal: PLoS ONE

    Article Title: ERK1/2 Signaling Plays an Important Role in Topoisomerase II Poison-Induced G2/M Checkpoint Activation

    doi: 10.1371/journal.pone.0050281

    Figure Lengend Snippet: DOX-induced G2/M checkpoint activation does not involve the p38 and MK2 kinases. (A) MCF-7 cells were treated for 2 hr with or without 1 μM DOX in the presence or absence of 50 μM U0126. As a positive control for p38 activation, a cell sample was exposed to UV at 100 J/m 2 and incubated for 1 hr at 37°C. The resulting cells were analyzed for levels of phospho-p38 ( p-p38 ) and total p38 ( p38 ) by immunoblotting. (B) The cell samples obtained above were analyzed for levels of phospho-MK2 ( pMK2 ), total MK2 ( MK2 ), phospho-ERK1/2 ( pERK1/2 ) and total ERK1/2 ( ERK1/2 ) by immunoblotting. (C) MCF-7 cells were transfected with control ( Control ) or MK2 specific siRNA ( MK2 ) and incubated for 2 days at 37°C. Upper panel: levels of MK2 and Actin in the transfected cells were determined by immunoblotting. Lower panel: The transfected cells were treated with 1 μM DOX, incubated for 24 hr and analyzed for DNA content by FACS. Graph depicts percentage of cells with 4N -DNA content and represents the mean ± s.d of two separate experiments with duplicate samples.

    Article Snippet: Cell culture and drug treatment MCF-7 and T47D human breast cancer cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS).

    Techniques: Activation Assay, Positive Control, Incubation, Transfection, FACS

    DOX and ETOP induce G2/M arrest and ERK1/2 activation in MCF-7 breast cancer cells. (A) Log-phase MCF-7 cells were treated with DOX or ETOP at the indicated doses as described in Materials and Methods and incubated for 24 hr. The cells were analyzed for DNA content by FACS. Upper panel: histograms shown are cells treated with none, 1 µM DOX or 10 µM ETOP. Cell cycle phases are indicated. Lower panel: Graphs depict the percentage of cells with 4 N -DNA content, indicative of G2/M phase of the cell cycle, and represent the mean ± s.d. of two sets of experiment with duplicate samples. (B) MCF-7 cells were incubated in the presence of 0.5 µM DOX or 10 µM ETOP for the hours indicated and analyzed for phospho-ERK1/2 and total-ERK1/2 by immunoblotting.

    Journal: PLoS ONE

    Article Title: ERK1/2 Signaling Plays an Important Role in Topoisomerase II Poison-Induced G2/M Checkpoint Activation

    doi: 10.1371/journal.pone.0050281

    Figure Lengend Snippet: DOX and ETOP induce G2/M arrest and ERK1/2 activation in MCF-7 breast cancer cells. (A) Log-phase MCF-7 cells were treated with DOX or ETOP at the indicated doses as described in Materials and Methods and incubated for 24 hr. The cells were analyzed for DNA content by FACS. Upper panel: histograms shown are cells treated with none, 1 µM DOX or 10 µM ETOP. Cell cycle phases are indicated. Lower panel: Graphs depict the percentage of cells with 4 N -DNA content, indicative of G2/M phase of the cell cycle, and represent the mean ± s.d. of two sets of experiment with duplicate samples. (B) MCF-7 cells were incubated in the presence of 0.5 µM DOX or 10 µM ETOP for the hours indicated and analyzed for phospho-ERK1/2 and total-ERK1/2 by immunoblotting.

    Article Snippet: Cell culture and drug treatment MCF-7 and T47D human breast cancer cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS).

    Techniques: Activation Assay, Incubation, FACS

    Effect of ERK1/2 inhibition on topo II poison-induced ATR and ATM signaling activation. (A) MCF-7 cells were treated for 2 hr with 1 µM DOX (upper panel) or 10 µM ETOP (lower panel) in the presence or absence of 50 µM U0126. The cells were washed and incubated in growth medium for additional 2 hr with the presence or absence of U0126. ATR and Chk1 kinase were respectively immunoprecipitated from the resulting cell lysates and assayed for kinase activity. ATR and Chk1 levels in immunoprecipitates were determined by immunoblotting ( ATR IP-WB and Chk1 IP-WB ). IgG , as a negative control, kinase assay was carried out using immunoprecipitates obtained by incubating control untreated cell lysate with non-immunized IgG. (B) Cells transfected with ERK1/2 specific or control siRNA were incubated for 2 days and treated with or without 0.5 µM DOX, as described above. ATR and Chk1 were respectively immunoprecipitated from the cell lysates and examined for kinase activity ( ATR activity and Chk1 activity ). ATR and Chk1 protein levels in immunoprecipitates were determined by immunoblotting ( ATR IP-WB and Chk1 IP-WB ). Levels of ERK1/2 and Actin in cell lysates were analyzed by immunoblotting ( ERK1/2 and Actin ). (C) Cells were treated as described in (A). ATM and Chk2 were immunoprecipitated from the cell lysates and assayed for kinase activity ( ATM activity and Chk2 activity ). ATM and Chk2 levels in immunoprecipitates were determined by immunoblotting ( ATM IP-WB and Chk2 IP-WB ). (D) MCF-7 cells were treated as described in (A) and incubated for the times indicated. Cdc2 was immunoprecipitated from cell lysate and analyzed for Cdc2-Tyr15 phosphorylation by immunoblotting ( Cdc2-Tyr15 ). Cdc2 in the immunoprecipitates was quantified by immunoblotting ( Cdc-2 IP-WB ).

    Journal: PLoS ONE

    Article Title: ERK1/2 Signaling Plays an Important Role in Topoisomerase II Poison-Induced G2/M Checkpoint Activation

    doi: 10.1371/journal.pone.0050281

    Figure Lengend Snippet: Effect of ERK1/2 inhibition on topo II poison-induced ATR and ATM signaling activation. (A) MCF-7 cells were treated for 2 hr with 1 µM DOX (upper panel) or 10 µM ETOP (lower panel) in the presence or absence of 50 µM U0126. The cells were washed and incubated in growth medium for additional 2 hr with the presence or absence of U0126. ATR and Chk1 kinase were respectively immunoprecipitated from the resulting cell lysates and assayed for kinase activity. ATR and Chk1 levels in immunoprecipitates were determined by immunoblotting ( ATR IP-WB and Chk1 IP-WB ). IgG , as a negative control, kinase assay was carried out using immunoprecipitates obtained by incubating control untreated cell lysate with non-immunized IgG. (B) Cells transfected with ERK1/2 specific or control siRNA were incubated for 2 days and treated with or without 0.5 µM DOX, as described above. ATR and Chk1 were respectively immunoprecipitated from the cell lysates and examined for kinase activity ( ATR activity and Chk1 activity ). ATR and Chk1 protein levels in immunoprecipitates were determined by immunoblotting ( ATR IP-WB and Chk1 IP-WB ). Levels of ERK1/2 and Actin in cell lysates were analyzed by immunoblotting ( ERK1/2 and Actin ). (C) Cells were treated as described in (A). ATM and Chk2 were immunoprecipitated from the cell lysates and assayed for kinase activity ( ATM activity and Chk2 activity ). ATM and Chk2 levels in immunoprecipitates were determined by immunoblotting ( ATM IP-WB and Chk2 IP-WB ). (D) MCF-7 cells were treated as described in (A) and incubated for the times indicated. Cdc2 was immunoprecipitated from cell lysate and analyzed for Cdc2-Tyr15 phosphorylation by immunoblotting ( Cdc2-Tyr15 ). Cdc2 in the immunoprecipitates was quantified by immunoblotting ( Cdc-2 IP-WB ).

    Article Snippet: Cell culture and drug treatment MCF-7 and T47D human breast cancer cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS).

    Techniques: Inhibition, Activation Assay, Incubation, Immunoprecipitation, Activity Assay, Western Blot, Negative Control, Kinase Assay, Transfection

    Effect of ATR and ATM expression on DOX-induced G2/M arrest. (A) MCF-7 cells stably expressing ATR specific shRNA ( ATR-shRNA ) or control shRNA ( Control-shRNA ) were treated with or without 0.5 µM DOX, incubated for 24 hr and analyzed for DNA content by FACS. Bar graph depicts the percentage of cells with 4N -DNA content and represents the mean ± s.d of two independent experiments with duplicate samples. * p

    Journal: PLoS ONE

    Article Title: ERK1/2 Signaling Plays an Important Role in Topoisomerase II Poison-Induced G2/M Checkpoint Activation

    doi: 10.1371/journal.pone.0050281

    Figure Lengend Snippet: Effect of ATR and ATM expression on DOX-induced G2/M arrest. (A) MCF-7 cells stably expressing ATR specific shRNA ( ATR-shRNA ) or control shRNA ( Control-shRNA ) were treated with or without 0.5 µM DOX, incubated for 24 hr and analyzed for DNA content by FACS. Bar graph depicts the percentage of cells with 4N -DNA content and represents the mean ± s.d of two independent experiments with duplicate samples. * p

    Article Snippet: Cell culture and drug treatment MCF-7 and T47D human breast cancer cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS).

    Techniques: Expressing, Stable Transfection, shRNA, Incubation, FACS

    Inhibition of ERK1/2 attenuates topo II poison-induced G2/M arrest in MCF-7 cells. (A) Upper panel: MCF-7 cells were incubated with U0126 at the indicated doses for 1 hr and then treated with 10 µM ETOP for 2 hr with the presence of U0126. The cells were lysed and analyzed for phospho- and total-ERK1/2. Lower panel: in the presence or absence of 50 µM U0126, MCF-7 cells were treated with ETOP at the indicated doses for 2 hr. The cells were washed, incubated for additional 24 hr in the presence or absence of U0126, and analyzed for DNA content by FACS. Bar graphs depict the percentage of cells with 4N -DNA content (G2/M phase cells) and represent the average of two independent experiments in duplicate. * p

    Journal: PLoS ONE

    Article Title: ERK1/2 Signaling Plays an Important Role in Topoisomerase II Poison-Induced G2/M Checkpoint Activation

    doi: 10.1371/journal.pone.0050281

    Figure Lengend Snippet: Inhibition of ERK1/2 attenuates topo II poison-induced G2/M arrest in MCF-7 cells. (A) Upper panel: MCF-7 cells were incubated with U0126 at the indicated doses for 1 hr and then treated with 10 µM ETOP for 2 hr with the presence of U0126. The cells were lysed and analyzed for phospho- and total-ERK1/2. Lower panel: in the presence or absence of 50 µM U0126, MCF-7 cells were treated with ETOP at the indicated doses for 2 hr. The cells were washed, incubated for additional 24 hr in the presence or absence of U0126, and analyzed for DNA content by FACS. Bar graphs depict the percentage of cells with 4N -DNA content (G2/M phase cells) and represent the average of two independent experiments in duplicate. * p

    Article Snippet: Cell culture and drug treatment MCF-7 and T47D human breast cancer cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS).

    Techniques: Inhibition, Incubation, FACS

    Effect of p53 expression on DOX-induced G2/M arrest. (A) Upper panel: MCF-7 cells stably expressing HPV-E6 ( E6 ) and control cells ( Control ) were treated with 1 µM DOX for the indicated hours and analyzed for levels of p53 and Actin by immunoblotting. Lower panel: HPV-E6 expressing and control cells were treated with or without 1 µM DOX in the presence or absence of 50 µM U0126, incubated for 24 hr and analyzed for DNA content by FACS. Bar graphs depict the percentage of cells with 4N -DNA content (G2/M phase cells) and represent the mean ± s.d of two independent experiments in duplicate. (B) MCF-7 cells were transfected with either vector expressing p53-R175H ( R175H ) dominant negative mutant or control empty vector ( Control ) and incubated for 48 hr. Upper panel: levels of p53 and Actin in the transfected or unstransfected cells were compared by immunoblotting. Lower panel: p53-R175H transfected and control cells were treated with 0.25 µM DOX in the presence or absence of U0126, incubated for 24 hr and analyzed for DNA content by FACS. Bar graphs depict the percentage of cells with 4N -DNA content and represent the mean ± s.d. of two independent experiments in duplicate. * p = 0.001 (n = 4), significant difference from the control transfected cells treated with DOX in the presence of U0126.

    Journal: PLoS ONE

    Article Title: ERK1/2 Signaling Plays an Important Role in Topoisomerase II Poison-Induced G2/M Checkpoint Activation

    doi: 10.1371/journal.pone.0050281

    Figure Lengend Snippet: Effect of p53 expression on DOX-induced G2/M arrest. (A) Upper panel: MCF-7 cells stably expressing HPV-E6 ( E6 ) and control cells ( Control ) were treated with 1 µM DOX for the indicated hours and analyzed for levels of p53 and Actin by immunoblotting. Lower panel: HPV-E6 expressing and control cells were treated with or without 1 µM DOX in the presence or absence of 50 µM U0126, incubated for 24 hr and analyzed for DNA content by FACS. Bar graphs depict the percentage of cells with 4N -DNA content (G2/M phase cells) and represent the mean ± s.d of two independent experiments in duplicate. (B) MCF-7 cells were transfected with either vector expressing p53-R175H ( R175H ) dominant negative mutant or control empty vector ( Control ) and incubated for 48 hr. Upper panel: levels of p53 and Actin in the transfected or unstransfected cells were compared by immunoblotting. Lower panel: p53-R175H transfected and control cells were treated with 0.25 µM DOX in the presence or absence of U0126, incubated for 24 hr and analyzed for DNA content by FACS. Bar graphs depict the percentage of cells with 4N -DNA content and represent the mean ± s.d. of two independent experiments in duplicate. * p = 0.001 (n = 4), significant difference from the control transfected cells treated with DOX in the presence of U0126.

    Article Snippet: Cell culture and drug treatment MCF-7 and T47D human breast cancer cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS).

    Techniques: Expressing, Stable Transfection, Incubation, FACS, Transfection, Plasmid Preparation, Dominant Negative Mutation

    Inhibition of ERK1/2 by specific siRNA diminishes topo II poison-induced G2/M arrest. (A) MCF-7 cells were transfected with ERK1/2 specific siRNA or control non-targeting siRNA and incubated for 2 days. The cells were then treated with 0.5 µM DOX or 5 µM ETOP, incubated for 24 hr and analyzed for DNA content by FACS. Left panel: histograms shown are DNA content analyses for the indicated cell samples. Upper right panel: levels of ERK1/2 in siRNA-transfected cells were determined by Western blotting. Lower right panel: bar graph depicts the percentage of cells in G2/M phase and presented as mean ± s.d. of three independent experiments in duplicate. ** p

    Journal: PLoS ONE

    Article Title: ERK1/2 Signaling Plays an Important Role in Topoisomerase II Poison-Induced G2/M Checkpoint Activation

    doi: 10.1371/journal.pone.0050281

    Figure Lengend Snippet: Inhibition of ERK1/2 by specific siRNA diminishes topo II poison-induced G2/M arrest. (A) MCF-7 cells were transfected with ERK1/2 specific siRNA or control non-targeting siRNA and incubated for 2 days. The cells were then treated with 0.5 µM DOX or 5 µM ETOP, incubated for 24 hr and analyzed for DNA content by FACS. Left panel: histograms shown are DNA content analyses for the indicated cell samples. Upper right panel: levels of ERK1/2 in siRNA-transfected cells were determined by Western blotting. Lower right panel: bar graph depicts the percentage of cells in G2/M phase and presented as mean ± s.d. of three independent experiments in duplicate. ** p

    Article Snippet: Cell culture and drug treatment MCF-7 and T47D human breast cancer cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS).

    Techniques: Inhibition, Transfection, Incubation, FACS, Western Blot