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  • 96
    New England Biolabs mfe
    Synthesis of plasmids containing aph (7″) or aac (3)IV as non-polar hygromycin B and apramycin resistance markers. (A) Schematic of ultramers designed to amplify aph (7″) or aac (3)IV. The 5′ ultramers 5631 and 5633, for aph (7″) or aac (3)IV respectively, include <t>Mfe</t> I, Kpn I and Sma I restriction sites, stop codons in all three reading frames, and a ribosome binding site. The 3′ ultramers 5632 and 5634 include a ribosome binding site, a start codon in-frame with restriction sites for Sma I and Bam HI, and restriction sites for Xba I, Nde I, Pst I and Sph I. (B) The amplified aac (3)IV was introduced by TA cloning into linearized pGEM-T, conserving the restriction sites in the pGEM-T multiple cloning site (MCS). The resulting plasmid is pAC1A. The pGEM sites may also be used for the sub-cloning of the apramycin resistance marker ( Apr R ). MCS sites that cut aac (3)IV are indicated with a superscript ‘A’. (C) All introduced sites in pAC1A were tested by restriction digest. (D) The Mfe I- and Sph I-digested aph (7″) amplification product was cloned into pBAD24 digested with Eco RI ( Mfe I-compatible) and Sph I. The <t>Mfe</t> I site was lost in the resulting plasmid, pAC1H. (E) All restriction sites introduced to pAC1H were tested by digest.
    Mfe, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Millipore mfe 280
    Differential expression of EMT/MET-related genes in <t>EAC-miR-200</t> depleted and UCS-miR-200 overexpressed cells. ( A ) Heatmap of RNA expression for 26 EMT/MET-related genes. Relative RNA expression of miR-200b/c-depleted vs scramble-treated Ishikawa and <t>MFE-280</t> cells, and miR-200c-overexpressed vs non-targeting control treated SNU-685 and JHUCS-1 cells, are shown. Increased and decreased expression are depicted in red and blue, respectively. The top 17 genes are known to have increased expression in the mesenchymal phenotype, while the bottom 9 genes are known to have increased expression in the epithelial phenotype. Relative expression is calculated by log2 fold-change ( B ).
    Mfe 280, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Millipore mfe 296
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Mfe 296, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher mfe
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Mfe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Eli Lilly mfe
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Mfe, supplied by Eli Lilly, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mfe  (Cerner)
    88
    Cerner mfe
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Mfe, supplied by Cerner, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Johnson & Johnson mfes
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Mfes, supplied by Johnson & Johnson, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Trion Research mfe i sal i
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Mfe I Sal I, supplied by Trion Research, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore nde i mfe
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Nde I Mfe, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher mfe i
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Mfe I, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    DSMZ mfe 296
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Mfe 296, supplied by DSMZ, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Sekisui XenoTech mfe supplement a
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Mfe Supplement A, supplied by Sekisui XenoTech, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Addgene inc mfe i digested pcag gfp
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Mfe I Digested Pcag Gfp, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Agilent technologies molecular feature extractor mfe algorithm
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Molecular Feature Extractor Mfe Algorithm, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 89/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Sekisui XenoTech mfe support medium
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Mfe Support Medium, supplied by Sekisui XenoTech, used in various techniques. Bioz Stars score: 88/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies bioconfirm mfe software
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Bioconfirm Mfe Software, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Agilent technologies molecular feature extraction mfe
    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) <t>MFE-296</t> and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.
    Molecular Feature Extraction Mfe, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 86/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Synthesis of plasmids containing aph (7″) or aac (3)IV as non-polar hygromycin B and apramycin resistance markers. (A) Schematic of ultramers designed to amplify aph (7″) or aac (3)IV. The 5′ ultramers 5631 and 5633, for aph (7″) or aac (3)IV respectively, include Mfe I, Kpn I and Sma I restriction sites, stop codons in all three reading frames, and a ribosome binding site. The 3′ ultramers 5632 and 5634 include a ribosome binding site, a start codon in-frame with restriction sites for Sma I and Bam HI, and restriction sites for Xba I, Nde I, Pst I and Sph I. (B) The amplified aac (3)IV was introduced by TA cloning into linearized pGEM-T, conserving the restriction sites in the pGEM-T multiple cloning site (MCS). The resulting plasmid is pAC1A. The pGEM sites may also be used for the sub-cloning of the apramycin resistance marker ( Apr R ). MCS sites that cut aac (3)IV are indicated with a superscript ‘A’. (C) All introduced sites in pAC1A were tested by restriction digest. (D) The Mfe I- and Sph I-digested aph (7″) amplification product was cloned into pBAD24 digested with Eco RI ( Mfe I-compatible) and Sph I. The Mfe I site was lost in the resulting plasmid, pAC1H. (E) All restriction sites introduced to pAC1H were tested by digest.

    Journal: PLoS ONE

    Article Title: Hygromycin B and Apramycin Antibiotic Resistance Cassettes for Use in Campylobacter jejuni

    doi: 10.1371/journal.pone.0095084

    Figure Lengend Snippet: Synthesis of plasmids containing aph (7″) or aac (3)IV as non-polar hygromycin B and apramycin resistance markers. (A) Schematic of ultramers designed to amplify aph (7″) or aac (3)IV. The 5′ ultramers 5631 and 5633, for aph (7″) or aac (3)IV respectively, include Mfe I, Kpn I and Sma I restriction sites, stop codons in all three reading frames, and a ribosome binding site. The 3′ ultramers 5632 and 5634 include a ribosome binding site, a start codon in-frame with restriction sites for Sma I and Bam HI, and restriction sites for Xba I, Nde I, Pst I and Sph I. (B) The amplified aac (3)IV was introduced by TA cloning into linearized pGEM-T, conserving the restriction sites in the pGEM-T multiple cloning site (MCS). The resulting plasmid is pAC1A. The pGEM sites may also be used for the sub-cloning of the apramycin resistance marker ( Apr R ). MCS sites that cut aac (3)IV are indicated with a superscript ‘A’. (C) All introduced sites in pAC1A were tested by restriction digest. (D) The Mfe I- and Sph I-digested aph (7″) amplification product was cloned into pBAD24 digested with Eco RI ( Mfe I-compatible) and Sph I. The Mfe I site was lost in the resulting plasmid, pAC1H. (E) All restriction sites introduced to pAC1H were tested by digest.

    Article Snippet: The initial aph (7″) PCR product was instead digested with Mfe I and Sph I (NEB), purified, and ligated to low-copy pBAD24 digested with Eco RI and Sph I.

    Techniques: Binding Assay, Amplification, TA Cloning, Clone Assay, Plasmid Preparation, Subcloning, Marker

    Differential expression of EMT/MET-related genes in EAC-miR-200 depleted and UCS-miR-200 overexpressed cells. ( A ) Heatmap of RNA expression for 26 EMT/MET-related genes. Relative RNA expression of miR-200b/c-depleted vs scramble-treated Ishikawa and MFE-280 cells, and miR-200c-overexpressed vs non-targeting control treated SNU-685 and JHUCS-1 cells, are shown. Increased and decreased expression are depicted in red and blue, respectively. The top 17 genes are known to have increased expression in the mesenchymal phenotype, while the bottom 9 genes are known to have increased expression in the epithelial phenotype. Relative expression is calculated by log2 fold-change ( B ).

    Journal: Scientific Reports

    Article Title: miR-200c-driven Mesenchymal-To-Epithelial Transition is a Therapeutic Target in Uterine Carcinosarcomas

    doi: 10.1038/s41598-017-03972-7

    Figure Lengend Snippet: Differential expression of EMT/MET-related genes in EAC-miR-200 depleted and UCS-miR-200 overexpressed cells. ( A ) Heatmap of RNA expression for 26 EMT/MET-related genes. Relative RNA expression of miR-200b/c-depleted vs scramble-treated Ishikawa and MFE-280 cells, and miR-200c-overexpressed vs non-targeting control treated SNU-685 and JHUCS-1 cells, are shown. Increased and decreased expression are depicted in red and blue, respectively. The top 17 genes are known to have increased expression in the mesenchymal phenotype, while the bottom 9 genes are known to have increased expression in the epithelial phenotype. Relative expression is calculated by log2 fold-change ( B ).

    Article Snippet: Cell culture Ishikawa and MFE-280 EAC cell lines were obtained from Sigma Aldrich; the HEC-251 EAC cell line was obtained from Riken BRC Cell Bank (Japan).

    Techniques: Expressing, RNA Expression

    Effects of consecutive transient miR-200 depletion in EAC cell lines. ( A ) Constitutive miR-200b/c depletion in EAC cell lines. Ishikawa and MFE-280 cell lines were transiently transfected with miR-200b/c inhibitor every 4 days for a total of 40 days (D = day, C = transfection cycle). Bar graphs represent relative miR-200b/c expression. Data are normalized to scramble. ( B ) Relative mRNA expression of EMT markers. TaqMan probes for ZEB1, ZEB2, E-cadherin (E-cad), N-cadherin (N-cad) and vimentin (vim) are shown on the bottom. Scramble (Scr) represents control for miR200b/c KD (200KD) samples. Data are normalized to scramble. Representative time points D12C3, D38C8 and D40C10 are shown. ( C ) Protein expression of EMT markers by immunoblotting. Representative early and late time points are shown. Anti-β-actin serves as a loading control. ( D ) Relative adhesion, and ( E ) migration and invasion in miR-200b/c knockdown (miR200b/c KD) vs. scramble-treated cells. ( F ) Relative proliferation measured by cell counts. Data are normalized to Day 0. All graphical data are reported as mean ± SD. *P

    Journal: Scientific Reports

    Article Title: miR-200c-driven Mesenchymal-To-Epithelial Transition is a Therapeutic Target in Uterine Carcinosarcomas

    doi: 10.1038/s41598-017-03972-7

    Figure Lengend Snippet: Effects of consecutive transient miR-200 depletion in EAC cell lines. ( A ) Constitutive miR-200b/c depletion in EAC cell lines. Ishikawa and MFE-280 cell lines were transiently transfected with miR-200b/c inhibitor every 4 days for a total of 40 days (D = day, C = transfection cycle). Bar graphs represent relative miR-200b/c expression. Data are normalized to scramble. ( B ) Relative mRNA expression of EMT markers. TaqMan probes for ZEB1, ZEB2, E-cadherin (E-cad), N-cadherin (N-cad) and vimentin (vim) are shown on the bottom. Scramble (Scr) represents control for miR200b/c KD (200KD) samples. Data are normalized to scramble. Representative time points D12C3, D38C8 and D40C10 are shown. ( C ) Protein expression of EMT markers by immunoblotting. Representative early and late time points are shown. Anti-β-actin serves as a loading control. ( D ) Relative adhesion, and ( E ) migration and invasion in miR-200b/c knockdown (miR200b/c KD) vs. scramble-treated cells. ( F ) Relative proliferation measured by cell counts. Data are normalized to Day 0. All graphical data are reported as mean ± SD. *P

    Article Snippet: Cell culture Ishikawa and MFE-280 EAC cell lines were obtained from Sigma Aldrich; the HEC-251 EAC cell line was obtained from Riken BRC Cell Bank (Japan).

    Techniques: Transfection, Expressing, Migration

    AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) MFE-296 and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.

    Journal: Cancer Biology & Therapy

    Article Title: Antitumor effects and molecular mechanisms of ponatinib on endometrial cancer cells harboring activating FGFR2 mutations

    doi: 10.1080/15384047.2015.1108492

    Figure Lengend Snippet: AP24534 inhibits the cell proliferation and FGFR2 kinase activity in endometrial cancer cells with activating FGFR2 mutations. (A) Cells were treated with indicated concentration of AP24534 for 48 h, and the cell viability was determined by the MTT reduction assay. We determined the value of IC 50 of each compound. The concentration of chemicals producing 50% inhibition of cell growth (IC 50 ) was estimated from semilog concentration-inhibition curves constructed using Prism software (GraphPad Software Inc..) (B) MFE-296 and AN3CA cells were treated with 0.1 μM of AP24534 for 48 h. The DNA content (propidium iodide) and cell cycle distribution was analyzed by flow cytometry as described in Material and Methods.

    Article Snippet: The invasion of MFE-296 and AN3CA cells was measured by using the Transwell chambers (Chemicon, Millipore, CA) according to the manufacturer's protocol.

    Techniques: Activity Assay, Concentration Assay, MTT Assay, Inhibition, Construct, Software, Flow Cytometry, Cytometry

    AP24534 decreases the phosphorylation of ERK and/or Akt in endometrial cancer cells. ( A , C ) MFE-296 cells and MFE280 cells were treated with indicated concentration of AP24534 and PD17307 for 48 h. Expression levels of both phosphorylated and total Akt and ERK1/2 were measured by Western blot analysis. PTEN expression was determined under the same experimental condition. ( B ) AN3CA cells were treated with AP24534 and PD173074 of the indicated concentrations for 48 h, and then activation/expression of Akt and ERK1/2 were examined by performing western blotting. ( D ) MFE-296 and AN3CA cells were treated with AP24534 of the indicated concentrations for 48 h, and then phosphorylation of PLCγ, PKCα, and STAT5 were assessed by immunoblot analysis. ( E ) In addition, phospho-level of PLCγ, and PKCα was analyzed in MFE280 under the same experimental condition.

    Journal: Cancer Biology & Therapy

    Article Title: Antitumor effects and molecular mechanisms of ponatinib on endometrial cancer cells harboring activating FGFR2 mutations

    doi: 10.1080/15384047.2015.1108492

    Figure Lengend Snippet: AP24534 decreases the phosphorylation of ERK and/or Akt in endometrial cancer cells. ( A , C ) MFE-296 cells and MFE280 cells were treated with indicated concentration of AP24534 and PD17307 for 48 h. Expression levels of both phosphorylated and total Akt and ERK1/2 were measured by Western blot analysis. PTEN expression was determined under the same experimental condition. ( B ) AN3CA cells were treated with AP24534 and PD173074 of the indicated concentrations for 48 h, and then activation/expression of Akt and ERK1/2 were examined by performing western blotting. ( D ) MFE-296 and AN3CA cells were treated with AP24534 of the indicated concentrations for 48 h, and then phosphorylation of PLCγ, PKCα, and STAT5 were assessed by immunoblot analysis. ( E ) In addition, phospho-level of PLCγ, and PKCα was analyzed in MFE280 under the same experimental condition.

    Article Snippet: The invasion of MFE-296 and AN3CA cells was measured by using the Transwell chambers (Chemicon, Millipore, CA) according to the manufacturer's protocol.

    Techniques: Concentration Assay, Expressing, Western Blot, Activation Assay

    AP24534 inhibits the anchorage-independent growth of endometrial cancer cells. ( A ) MFE-296 cells were treated with AP24534 (0.01 and 0.1 μM) and PD 173074 (0.1 and 1 μM) twice a week for 3 wks in the soft-agar colony formation assay. ( B ) AN3CA cells were treated with AP24534 (0.001 and 0.01 μM) and PD173074 (0.01 and 0.1 μM) as described in Material and Methods. Colonies were counted using a digital microscope (Olympus DP71, USA) Colony counts from different treatment groups were subjected to statistical analysis. *, p

    Journal: Cancer Biology & Therapy

    Article Title: Antitumor effects and molecular mechanisms of ponatinib on endometrial cancer cells harboring activating FGFR2 mutations

    doi: 10.1080/15384047.2015.1108492

    Figure Lengend Snippet: AP24534 inhibits the anchorage-independent growth of endometrial cancer cells. ( A ) MFE-296 cells were treated with AP24534 (0.01 and 0.1 μM) and PD 173074 (0.1 and 1 μM) twice a week for 3 wks in the soft-agar colony formation assay. ( B ) AN3CA cells were treated with AP24534 (0.001 and 0.01 μM) and PD173074 (0.01 and 0.1 μM) as described in Material and Methods. Colonies were counted using a digital microscope (Olympus DP71, USA) Colony counts from different treatment groups were subjected to statistical analysis. *, p

    Article Snippet: The invasion of MFE-296 and AN3CA cells was measured by using the Transwell chambers (Chemicon, Millipore, CA) according to the manufacturer's protocol.

    Techniques: Soft Agar Assay, Microscopy

    AP24534 inhibits the migration and invasion of endometrial cancer cells. ( A ) MFE-296 and AN3CA cells were treated with indicated concentration of AP24534 and PD173074. Cell migration was measured by using the Culture-Inserts, and wound closure was monitored by photography at 36 h following treatment with each compound. ( B ) After treatment of MFE-296 and AN3CA cells with AP24534 (1 μM) and PD173074 (1 μM) for 48 h, an in vitro invasion assay was performed using a 24-well Transwell unit with polycarbonate filters having a diameter of 6.5 mm and a pore size of 8.0 μm, and the number of invading cells per field was counted under light microscopy. *, significantly different between the groups compared ( p

    Journal: Cancer Biology & Therapy

    Article Title: Antitumor effects and molecular mechanisms of ponatinib on endometrial cancer cells harboring activating FGFR2 mutations

    doi: 10.1080/15384047.2015.1108492

    Figure Lengend Snippet: AP24534 inhibits the migration and invasion of endometrial cancer cells. ( A ) MFE-296 and AN3CA cells were treated with indicated concentration of AP24534 and PD173074. Cell migration was measured by using the Culture-Inserts, and wound closure was monitored by photography at 36 h following treatment with each compound. ( B ) After treatment of MFE-296 and AN3CA cells with AP24534 (1 μM) and PD173074 (1 μM) for 48 h, an in vitro invasion assay was performed using a 24-well Transwell unit with polycarbonate filters having a diameter of 6.5 mm and a pore size of 8.0 μm, and the number of invading cells per field was counted under light microscopy. *, significantly different between the groups compared ( p

    Article Snippet: The invasion of MFE-296 and AN3CA cells was measured by using the Transwell chambers (Chemicon, Millipore, CA) according to the manufacturer's protocol.

    Techniques: Migration, Concentration Assay, In Vitro, Invasion Assay, Light Microscopy

    (A) MFE-296 cells were treated with the indicated concentrations of AP24534 (0.1 and 1 μM) or PD173074 (1 μM). FGFR2 was immunoprecipitated with FGFR2 antibody and FGFR2 kinase assay was performed as describe in Materials Methods section. *, p

    Journal: Cancer Biology & Therapy

    Article Title: Antitumor effects and molecular mechanisms of ponatinib on endometrial cancer cells harboring activating FGFR2 mutations

    doi: 10.1080/15384047.2015.1108492

    Figure Lengend Snippet: (A) MFE-296 cells were treated with the indicated concentrations of AP24534 (0.1 and 1 μM) or PD173074 (1 μM). FGFR2 was immunoprecipitated with FGFR2 antibody and FGFR2 kinase assay was performed as describe in Materials Methods section. *, p

    Article Snippet: The invasion of MFE-296 and AN3CA cells was measured by using the Transwell chambers (Chemicon, Millipore, CA) according to the manufacturer's protocol.

    Techniques: Immunoprecipitation, Kinase Assay

    Generation of sample mixes for analytic validation DNA from seven original samples was diluted to generate seven mixes. A. AN3CA and MFE-296 cell lines were mixed 1:1 to create Mix A, which was then mixed 1:1 with HCC827 to create Mix B and once again to create Mix C. B. Two frozen tumor samples and C. two FFPE tumor samples were combined 1:1 and 85:15 to generate mixes D and E (frozen) and F and G (FFPE).

    Journal: Oncotarget

    Article Title: Analytic validation and real-time clinical application of an amplicon-based targeted gene panel for advanced cancer

    doi: 10.18632/oncotarget.20616

    Figure Lengend Snippet: Generation of sample mixes for analytic validation DNA from seven original samples was diluted to generate seven mixes. A. AN3CA and MFE-296 cell lines were mixed 1:1 to create Mix A, which was then mixed 1:1 with HCC827 to create Mix B and once again to create Mix C. B. Two frozen tumor samples and C. two FFPE tumor samples were combined 1:1 and 85:15 to generate mixes D and E (frozen) and F and G (FFPE).

    Article Snippet: MFE-296 cells (Sigma Aldrich) were cultured in minimal essential media (MEM) (Sigma Aldrich) supplemented with 2 mM L-Glutamine (Sigma Aldrich) and 10% Fetal Bovine Serum (Sigma Aldrich).

    Techniques: Formalin-fixed Paraffin-Embedded

    FOXA1 promotes proliferation of human EC cells by affecting AR-mediated transcription. A : Proliferation in MFE-296 cells transfected with NC or shFOXA1 was assessed by the colony-forming assay (Left) and further quantified in the number of colonies of triplicate experiments (Right). B : Proliferation in AN3CA cells transfected with NC or exFOXA1 was assessed by the colony-forming assay (Left) and further quantified in the number of colonies of triplicate experiments (Right). C : Assessment of proliferation by the MTT assay in MFE-296 cells transfected with NC or shFOXA1. D : Assessment of proliferation by the MTT assay in AN3CA cells transfected with NC or exFOXA1. E : Left: Colony-formation assay of untransfected MFE-296 cells (WT) and MFE-296 cells transfected with NC, shFOXA1, or shFOXA1 and exAR. Right: Graphical representation of the fold change in the number of colonies in untransfected MFE-296 cells (MFE-296) and MFE-296 cells transfected with NC (MFE-296/NC), shFOXA1 (MFE-296/shFOXA1), or shFOXA1 and exAR (MFE-296/shFOXA1 + exAR). F : Proliferation of MFE-296, MFE-296/NC, MFE-296/shFOXA1, or MFE-296/shFOXA1 + exAR cells was assessed by MTT assay. The right panel reiterates the data in the left panel at 72 h. *p

    Journal: BMC Cancer

    Article Title: FOXA1 promotes tumor cell proliferation through AR involving the Notch pathway in endometrial cancer

    doi: 10.1186/1471-2407-14-78

    Figure Lengend Snippet: FOXA1 promotes proliferation of human EC cells by affecting AR-mediated transcription. A : Proliferation in MFE-296 cells transfected with NC or shFOXA1 was assessed by the colony-forming assay (Left) and further quantified in the number of colonies of triplicate experiments (Right). B : Proliferation in AN3CA cells transfected with NC or exFOXA1 was assessed by the colony-forming assay (Left) and further quantified in the number of colonies of triplicate experiments (Right). C : Assessment of proliferation by the MTT assay in MFE-296 cells transfected with NC or shFOXA1. D : Assessment of proliferation by the MTT assay in AN3CA cells transfected with NC or exFOXA1. E : Left: Colony-formation assay of untransfected MFE-296 cells (WT) and MFE-296 cells transfected with NC, shFOXA1, or shFOXA1 and exAR. Right: Graphical representation of the fold change in the number of colonies in untransfected MFE-296 cells (MFE-296) and MFE-296 cells transfected with NC (MFE-296/NC), shFOXA1 (MFE-296/shFOXA1), or shFOXA1 and exAR (MFE-296/shFOXA1 + exAR). F : Proliferation of MFE-296, MFE-296/NC, MFE-296/shFOXA1, or MFE-296/shFOXA1 + exAR cells was assessed by MTT assay. The right panel reiterates the data in the left panel at 72 h. *p

    Article Snippet: The human endometrial cell line MFE-296 was purchased from Sigma (St. Louis, MO, USA).

    Techniques: Transfection, MTT Assay, Colony Assay

    Tumorigenicity assay in nude mice. A : The growth rates of tumors formed from untransfected MFE-296 cells (MFE-296) and MFE-296 cells transfected with NC (MFE-296/NC) or shFOXA1 (MFE-296/shFOXA1). After injection, tumor volumes were calculated every seven days. B and C : Six weeks after injection of MFE-296, MFE-296-NC, and MFE-296-shFOXA1 cells, tumors were removed, and the tumor weights and volumes were determined. Arithmetic means and SD are shown. D : Staining with hematoxylin and eosin (H E) or immunohistochemical staining for FOXA1, AR, Notch1, Hes1, Ki67, and PCNA in mouse tumor tissues (immunohistochemical staining, 200×). *p

    Journal: BMC Cancer

    Article Title: FOXA1 promotes tumor cell proliferation through AR involving the Notch pathway in endometrial cancer

    doi: 10.1186/1471-2407-14-78

    Figure Lengend Snippet: Tumorigenicity assay in nude mice. A : The growth rates of tumors formed from untransfected MFE-296 cells (MFE-296) and MFE-296 cells transfected with NC (MFE-296/NC) or shFOXA1 (MFE-296/shFOXA1). After injection, tumor volumes were calculated every seven days. B and C : Six weeks after injection of MFE-296, MFE-296-NC, and MFE-296-shFOXA1 cells, tumors were removed, and the tumor weights and volumes were determined. Arithmetic means and SD are shown. D : Staining with hematoxylin and eosin (H E) or immunohistochemical staining for FOXA1, AR, Notch1, Hes1, Ki67, and PCNA in mouse tumor tissues (immunohistochemical staining, 200×). *p

    Article Snippet: The human endometrial cell line MFE-296 was purchased from Sigma (St. Louis, MO, USA).

    Techniques: Tumorigenicity Assay, Mouse Assay, Transfection, Injection, Staining, Immunohistochemistry

    FOXA1 affects the expression of AR in human EC cells. A : FOXA1 and AR expression in the indicated EC cell lines as determined were measured by western blotting (Left), and further quantified by densitometry of triplicate experiments (Right). β-actin was used as a loading control. B : Stable transfection of MFE-296 cells with negative control vector (MFE-296-NC) or shFOXA1 (MFE-296-shFOXA1). By comparing the cells in white light (the upper panels) with the cells in green fluorescence (the lower panels), the percentage of transfected/fluorescing cells was estimated at > 85%. Magnification, ×400. C : Quantification of FOXA1 mRNA by qRT-PCR in untransfected MFE-296 (MFE-296), MFE-296 transfected with shRNA control plasmid (MFE-296/NC), and MFE-296 transfected with shFOXA1 (MFE-296/shFOXA1). D : Quantification of AR mRNA by qRT-PCR in MFE-296, MFE-296/NC, and MFE-296/shFOXA1 cells. E : FOXA1 and AR expression in MFE-296, MFE-296/NC and MFE-296/shFOXA1 cells were measured by western blotting (Left), and further quantified by densitometry of triplicate experiments (Right). F : Quantification of FOXA1 mRNA by qRT-PCR in untransfected AN3CA (AN3CA), AN3CA transfected with control plasmid (AN3CA/NC), and AN3CA transfected with FOXA1 expression plasmid (AN3CA/exFOXA1). G : Quantification of AR mRNA by qRT-PCR in AN3CA, AN3CA/NC, and AN3CA/exFOXA1 cells. H : AR and FOXA1 expression in AN3CA, AN3CA/NC and AN3CA/exFOXA1 cells were measured by western blotting (Left), and further quantified by densitometry of triplicate experiments (Right). *p

    Journal: BMC Cancer

    Article Title: FOXA1 promotes tumor cell proliferation through AR involving the Notch pathway in endometrial cancer

    doi: 10.1186/1471-2407-14-78

    Figure Lengend Snippet: FOXA1 affects the expression of AR in human EC cells. A : FOXA1 and AR expression in the indicated EC cell lines as determined were measured by western blotting (Left), and further quantified by densitometry of triplicate experiments (Right). β-actin was used as a loading control. B : Stable transfection of MFE-296 cells with negative control vector (MFE-296-NC) or shFOXA1 (MFE-296-shFOXA1). By comparing the cells in white light (the upper panels) with the cells in green fluorescence (the lower panels), the percentage of transfected/fluorescing cells was estimated at > 85%. Magnification, ×400. C : Quantification of FOXA1 mRNA by qRT-PCR in untransfected MFE-296 (MFE-296), MFE-296 transfected with shRNA control plasmid (MFE-296/NC), and MFE-296 transfected with shFOXA1 (MFE-296/shFOXA1). D : Quantification of AR mRNA by qRT-PCR in MFE-296, MFE-296/NC, and MFE-296/shFOXA1 cells. E : FOXA1 and AR expression in MFE-296, MFE-296/NC and MFE-296/shFOXA1 cells were measured by western blotting (Left), and further quantified by densitometry of triplicate experiments (Right). F : Quantification of FOXA1 mRNA by qRT-PCR in untransfected AN3CA (AN3CA), AN3CA transfected with control plasmid (AN3CA/NC), and AN3CA transfected with FOXA1 expression plasmid (AN3CA/exFOXA1). G : Quantification of AR mRNA by qRT-PCR in AN3CA, AN3CA/NC, and AN3CA/exFOXA1 cells. H : AR and FOXA1 expression in AN3CA, AN3CA/NC and AN3CA/exFOXA1 cells were measured by western blotting (Left), and further quantified by densitometry of triplicate experiments (Right). *p

    Article Snippet: The human endometrial cell line MFE-296 was purchased from Sigma (St. Louis, MO, USA).

    Techniques: Expressing, Western Blot, Stable Transfection, Negative Control, Plasmid Preparation, Fluorescence, Transfection, Quantitative RT-PCR, shRNA

    FOXA1 affects AR-mediated transcription. A : MFE-296 cells were treated with DHT (10 −9 to 10 −7 M) or vehicle (control) for 24 h. qRT-PCR was used to assess the levels of AR, XBP1, MYC, ZBTB16, and UHRF1 mRNA. The levels of each mRNA are shown relative to the level expressed in the vehicle sample. B : Quantification of AR, XBP1, MYC, ZBTB16, and UHRF1 mRNA by qRT-PCR in MFE-296 cells treated with 10 −7 M DHT for 0–48 h. C : Western blotting analysis of AR in MFE-296 cells treated with vehicle, 10 −7 M DHT, or 10 −7 M DHT plus 10 −6 M flutamide (DHT + FLU) for 24 h. β-actin was used as a loading control. D : MFE-296/NC and MFE-296/shFOXA1 cells were treated with 10 −7 M DHT or vehicle for 24 h followed by qRT-PCR analysis of AR, XBP1, MYC, ZBTB16, and UHRF1 mRNA. E : AN3CA/NC and AN3CA/exFOXA1 cells were treated with 10 −7 M DHT or vehicle for 24 h followed by qRT-PCR analysis of AR, XBP1, MYC, ZBTB16, and UHRF1 mRNA. F : Quantification of AR expression by qRT-PCR and western blotting in untransfected MFE-296 cells (MFE-296) and MFE-296 cells transfected with NC (MFE-296/NC) or exAR (MFE-296/exAR). G : Expression of XBP1, MYC, ZBTB16, and UHRF1 mRNA in untransfected MFE-296 cells and MFE-296 cells transfected with NC, shFOXA1, or shFOXA1 and exAR was measured by qRT-PCR. H : Quantification of AR expression by qRT-PCR and western blotting in untransfected AN3CA cells (AN3CA) and AN3CA cells transfected with NC (AN3CA/NC) or siAR (AN3CA/siAR). I : Expression of XBP1, MYC, ZBTB16, and UHRF1 mRNA in untransfected AN3CA cells and AN3CA cells transfected with NC, exFOXA1, or exFOXA1 and siAR was measured by qRT-PCR. *p

    Journal: BMC Cancer

    Article Title: FOXA1 promotes tumor cell proliferation through AR involving the Notch pathway in endometrial cancer

    doi: 10.1186/1471-2407-14-78

    Figure Lengend Snippet: FOXA1 affects AR-mediated transcription. A : MFE-296 cells were treated with DHT (10 −9 to 10 −7 M) or vehicle (control) for 24 h. qRT-PCR was used to assess the levels of AR, XBP1, MYC, ZBTB16, and UHRF1 mRNA. The levels of each mRNA are shown relative to the level expressed in the vehicle sample. B : Quantification of AR, XBP1, MYC, ZBTB16, and UHRF1 mRNA by qRT-PCR in MFE-296 cells treated with 10 −7 M DHT for 0–48 h. C : Western blotting analysis of AR in MFE-296 cells treated with vehicle, 10 −7 M DHT, or 10 −7 M DHT plus 10 −6 M flutamide (DHT + FLU) for 24 h. β-actin was used as a loading control. D : MFE-296/NC and MFE-296/shFOXA1 cells were treated with 10 −7 M DHT or vehicle for 24 h followed by qRT-PCR analysis of AR, XBP1, MYC, ZBTB16, and UHRF1 mRNA. E : AN3CA/NC and AN3CA/exFOXA1 cells were treated with 10 −7 M DHT or vehicle for 24 h followed by qRT-PCR analysis of AR, XBP1, MYC, ZBTB16, and UHRF1 mRNA. F : Quantification of AR expression by qRT-PCR and western blotting in untransfected MFE-296 cells (MFE-296) and MFE-296 cells transfected with NC (MFE-296/NC) or exAR (MFE-296/exAR). G : Expression of XBP1, MYC, ZBTB16, and UHRF1 mRNA in untransfected MFE-296 cells and MFE-296 cells transfected with NC, shFOXA1, or shFOXA1 and exAR was measured by qRT-PCR. H : Quantification of AR expression by qRT-PCR and western blotting in untransfected AN3CA cells (AN3CA) and AN3CA cells transfected with NC (AN3CA/NC) or siAR (AN3CA/siAR). I : Expression of XBP1, MYC, ZBTB16, and UHRF1 mRNA in untransfected AN3CA cells and AN3CA cells transfected with NC, exFOXA1, or exFOXA1 and siAR was measured by qRT-PCR. *p

    Article Snippet: The human endometrial cell line MFE-296 was purchased from Sigma (St. Louis, MO, USA).

    Techniques: Quantitative RT-PCR, Western Blot, Expressing, Transfection

    FOXA1 affects AR-mediated transcription via binding with AR and activates the Notch pathway. A : Co-immunoprecipitation (IP) of FOXA1 with AR in MFE-296 cells. WB: western blotting. B : Co-immunoprecipitation of FOXA1 with AR in AN3CA cells treated with 10 −7 M DHT or vehicle. C : Schematic representation of the MYC locus. FOXA1-binding sites and AR-binding sites upstream of the TSS of MYC were predicted by ChIP-seq analysis. ChIP-PCR assays were performed using anti-FOXA1 antibody or anti-AR antibody. Pro: promoter region; Enh-1: enhancer 1 region; End-2: enhancer 2 region; TSS: transcription starting sites. D : Immunoprecipitated DNA fragments in ChIP-PCR assays were examined by qRT-PCR. Each sample was assayed in triplicate in each of three independent experiments. IgG was used as negative control. Primers were designed specifically for the promoter region (Pro), the enhancer 1 region (Enh-1), and the three putative FOXA1-AR binding sites within enhancer 2 region (Enh-2a, Enh-2b, and Enh-2c) according to the study [ 19 ]. E : Protein levels of FOXA1, AR, Notch1, and Hes1 in untransfected MFE-296 cells (MFE-296) and MFE-296 cells transfected with NC (MFE-296/NC), shFOXA1 (MFE-296/shFOXA1), or shFOXA1 and exAR (MFE-296/shFOXA1 + exAR) were measured by western blotting (Right), and further quantified by densitometry of triplicate experiments (Left). F : Protein levels of FOXA1, AR, Notch1, and Hes1 in untransfected AN3CA cells (AN3CA) and AN3CA cells transfected with NC (AN3CA/NC) , exFOXA1 (AN3CA/exFOXA1), or exFOXA1 and siAR (AN3CA/exFOXA1 + siAR) were measured by western blotting (Right), and further quantified by densitometry of triplicate experiments (Left). β-actin was used as a loading control. *p

    Journal: BMC Cancer

    Article Title: FOXA1 promotes tumor cell proliferation through AR involving the Notch pathway in endometrial cancer

    doi: 10.1186/1471-2407-14-78

    Figure Lengend Snippet: FOXA1 affects AR-mediated transcription via binding with AR and activates the Notch pathway. A : Co-immunoprecipitation (IP) of FOXA1 with AR in MFE-296 cells. WB: western blotting. B : Co-immunoprecipitation of FOXA1 with AR in AN3CA cells treated with 10 −7 M DHT or vehicle. C : Schematic representation of the MYC locus. FOXA1-binding sites and AR-binding sites upstream of the TSS of MYC were predicted by ChIP-seq analysis. ChIP-PCR assays were performed using anti-FOXA1 antibody or anti-AR antibody. Pro: promoter region; Enh-1: enhancer 1 region; End-2: enhancer 2 region; TSS: transcription starting sites. D : Immunoprecipitated DNA fragments in ChIP-PCR assays were examined by qRT-PCR. Each sample was assayed in triplicate in each of three independent experiments. IgG was used as negative control. Primers were designed specifically for the promoter region (Pro), the enhancer 1 region (Enh-1), and the three putative FOXA1-AR binding sites within enhancer 2 region (Enh-2a, Enh-2b, and Enh-2c) according to the study [ 19 ]. E : Protein levels of FOXA1, AR, Notch1, and Hes1 in untransfected MFE-296 cells (MFE-296) and MFE-296 cells transfected with NC (MFE-296/NC), shFOXA1 (MFE-296/shFOXA1), or shFOXA1 and exAR (MFE-296/shFOXA1 + exAR) were measured by western blotting (Right), and further quantified by densitometry of triplicate experiments (Left). F : Protein levels of FOXA1, AR, Notch1, and Hes1 in untransfected AN3CA cells (AN3CA) and AN3CA cells transfected with NC (AN3CA/NC) , exFOXA1 (AN3CA/exFOXA1), or exFOXA1 and siAR (AN3CA/exFOXA1 + siAR) were measured by western blotting (Right), and further quantified by densitometry of triplicate experiments (Left). β-actin was used as a loading control. *p

    Article Snippet: The human endometrial cell line MFE-296 was purchased from Sigma (St. Louis, MO, USA).

    Techniques: Binding Assay, Immunoprecipitation, Western Blot, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Quantitative RT-PCR, Negative Control, Transfection

    FOXA1 induces migration and invasion in EC cells. A : Cell migration of MFE-296, MFE-296/NC, MFE-296/shFOXA1 and MFE-296/shFOXA1 + exAR cells was assessed by the transwell migration analysis (Left). The mean ± SD number of migrated cells of three independent experiments was showed in the right panel. The abbreviation “HPF” on the y axis means one high power field. B : Cell migration of AN3CA, AN3CA/NC, AN3CA/exFOXA1 and AN3CA/exFOXA1 + siAR cells were subjected to transwell migration analysis (Left). The mean ± SD number of migrated cells of three independent experiments was showed in the right panel. C : Cell invasion of MFE-296, MFE-296/NC, MFE-296/shFOXA1 and MFE-296/shFOXA1 + exAR cells was assessed by the transwell invasion analysis (Left). The mean ± SD number of invased cells of three independent experiments was shown in the right panel. D : Cell invasion of AN3CA, AN3CA/NC, AN3CA/exFOXA1 and AN3CA/exFOXA1 + siAR cells by three independent experiments were subjected to transwell invasion analysis (Left). The mean ± SD number of invased cells of three independent experiments was showed in the right panel. (Magnification, 200×). *p

    Journal: BMC Cancer

    Article Title: FOXA1 promotes tumor cell proliferation through AR involving the Notch pathway in endometrial cancer

    doi: 10.1186/1471-2407-14-78

    Figure Lengend Snippet: FOXA1 induces migration and invasion in EC cells. A : Cell migration of MFE-296, MFE-296/NC, MFE-296/shFOXA1 and MFE-296/shFOXA1 + exAR cells was assessed by the transwell migration analysis (Left). The mean ± SD number of migrated cells of three independent experiments was showed in the right panel. The abbreviation “HPF” on the y axis means one high power field. B : Cell migration of AN3CA, AN3CA/NC, AN3CA/exFOXA1 and AN3CA/exFOXA1 + siAR cells were subjected to transwell migration analysis (Left). The mean ± SD number of migrated cells of three independent experiments was showed in the right panel. C : Cell invasion of MFE-296, MFE-296/NC, MFE-296/shFOXA1 and MFE-296/shFOXA1 + exAR cells was assessed by the transwell invasion analysis (Left). The mean ± SD number of invased cells of three independent experiments was shown in the right panel. D : Cell invasion of AN3CA, AN3CA/NC, AN3CA/exFOXA1 and AN3CA/exFOXA1 + siAR cells by three independent experiments were subjected to transwell invasion analysis (Left). The mean ± SD number of invased cells of three independent experiments was showed in the right panel. (Magnification, 200×). *p

    Article Snippet: The human endometrial cell line MFE-296 was purchased from Sigma (St. Louis, MO, USA).

    Techniques: Migration