antibodies against her2 Search Results


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  • 95
    Thermo Fisher antibodies against her2
    Western blots confirmed low (MCF7 parental) intermediate (Clone A) and high (Clone B) <t>Her2</t> expression. On the left (no 17-DMAG, dose 0), samples of 3 experiments were loaded into 1 gel to show the standard error of the mean. Her2 expression decreased
    Antibodies Against Her2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Millipore antibodies against her 2
    Figure 4A: Western blotting analysis of LTLT-Ca cells treated with entinostat alone or in presence of MG-132 (proteosomal inhibitor) or NH 4 Cl (lysosomal inhibitor): LTLT-Ca cells were treated with ENT (1μM) alone or in presence of MG-132 (5μM) or NH 4 Cl (100μM) or both. Protein expression in the cells was examined by western immunoblotting as described in “materials and methods”. Blots were probed for β-actin to verify equal loading. The numbers below the blots show densitometric values that are corrected for loading. Figure 4B: Western blotting analysis of SKBr3 and BT-474 cells treated with entinostat: <t>Her-2</t> positive SKBr3 and BT-474 cells were treated with ENT (1μM). Protein expression in the cells was examined by western immunoblotting as described in “materials and methods”. Blots were probed for β-actin to verify equal loading. The numbers below the blots show densitometric values that are corrected for loading. Figure 4C: Western blotting analysis of AnR and ExR cells treated with entinostat: Her-2 positive anastrozole resistant (AnR) and exemestane resistant (ExR) cells were treated with ENT (1μM). Protein expression in the cells was examined by western immunoblotting as described in “materials and methods”. Blots were probed for β-actin to verify equal loading. The numbers below the blots show densitometric values that are corrected for loading.
    Antibodies Against Her 2, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Cell Signaling Technology Inc antibodies against her2
    Let-7f regulates β2-AR expression in breast cancer cells. a to d , MCF-7 ( a and b ) and <t>MCF-7/Her2</t> cells ( c and d ) were planted in 24-well plates and transfected with 9 and 27 pmol synthetic inhibitors or mimics of let-7f. The expression of β2-AR was analyzed by Western blot. These experiments were repeated twice
    Antibodies Against Her2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 89/100, based on 88 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Agilent technologies antibody against her2
    Immunohistochemistry of <t>Her2</t> and GEP100 in adenocarcinoma lung tissue specimens. ( A ) Her2 and GEP100 were each stained in brown in sequential sections. Case 1, Her2 and GEP100 double negative; Case 2, Her2 score 1+ and GEP100 score 1+; Case 3, Her2 score 2+ and GEP100 score 2+; Case 4, Her2 score 3+ GEP100 and GEP100 score 2+. All sections were counterstained with hematoxylin and eosin (H E) and representative figures are shown. Scale bars, 100 µm. ( B ) Representative figures of the peripheral expression of GEP100. Scale bars, 500 µm. ( C ) No correlation between strong expression of GEP100 (score 2+) per se with node-metastases. ( D ) A statistical correlation between double strong positive signal of GEP100 (score 2+) and Her2 (score 3+), and node-metastases. In C and D , solid bars mean GEP100 positive rate for strong expression (score 2+).
    Antibody Against Her2, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 89/100, based on 114 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Abcam antibodies against her2
    (A) Expression of HER-2 mRNA, as detected by reverse transcription-quantitative polymerase chain reaction following transfection with the lentiviral-mediated <t>HER2-small</t> hairpin RNA. Differences in the expression of HER-2 mRNA between the KD and NC and CON groups were significant (P
    Antibodies Against Her2, supplied by Abcam, used in various techniques. Bioz Stars score: 89/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Agilent technologies polyclonal antibody against her2
    (A) Expression of HER-2 mRNA, as detected by reverse transcription-quantitative polymerase chain reaction following transfection with the lentiviral-mediated <t>HER2-small</t> hairpin RNA. Differences in the expression of HER-2 mRNA between the KD and NC and CON groups were significant (P
    Polyclonal Antibody Against Her2, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 85/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Santa Cruz Biotechnology antibodies against her2
    HBx increased <t>HER2</t> protein expression by HuR-dependent mRNA stabilization in HCC cells. (a) The HuR protein expression in HBx-paired HCC cells was examined by Western blot ( N = 4). (b) Hep3Bx cells were transiently transfected with either si-control or si-HuR for 4 days. The protein expressions of HER2, EGFR, and HuR were analyzed by Western blot ( N = 3). (c) Hep3Bx cells were transiently transfected with either si-control or si-HuR for 4 days, followed by the treatment of 5 μ M Actinomycin D. The relative remaining HER2 mRNA expression in each group was determined by RT-qPCR. The HER2 mRNA expression was normalized to actin expression. Statistical analysis was performed by Student's t -test. * P
    Antibodies Against Her2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 89/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Agilent technologies antibodies against her2 neu
    Detection of ovarian metastases by a combination of markers. Representative image of a lobular ovarian metastasis stained with DAPI counterstain and triple immunofluorescence for EpCAM ( a ), EMA ( b ), <t>Her2/neu</t> ( c ), and the three stainings combined ( d ). The solid arrow indicates tumor cells that are positive for EpCAM, but negative for EMA and Her2/neu. The dashed arrow indicates tumor cells that are positive for Her2/neu, but negative for EpCAM and EMA. Scale bars represent 100 μm. EpCAM, epithelial cell adhesion molecule; EMA, epithelial membrane antigen; Her2/neu, human epidermal growth receptor type 2
    Antibodies Against Her2 Neu, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 90/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies polyclonal antibody against her2 neu
    Detection of ovarian metastases by a combination of markers. Representative image of a lobular ovarian metastasis stained with DAPI counterstain and triple immunofluorescence for EpCAM ( a ), EMA ( b ), <t>Her2/neu</t> ( c ), and the three stainings combined ( d ). The solid arrow indicates tumor cells that are positive for EpCAM, but negative for EMA and Her2/neu. The dashed arrow indicates tumor cells that are positive for Her2/neu, but negative for EpCAM and EMA. Scale bars represent 100 μm. EpCAM, epithelial cell adhesion molecule; EMA, epithelial membrane antigen; Her2/neu, human epidermal growth receptor type 2
    Polyclonal Antibody Against Her2 Neu, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 85/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc antibodies against her2 erbb2
    Detection of ovarian metastases by a combination of markers. Representative image of a lobular ovarian metastasis stained with DAPI counterstain and triple immunofluorescence for EpCAM ( a ), EMA ( b ), <t>Her2/neu</t> ( c ), and the three stainings combined ( d ). The solid arrow indicates tumor cells that are positive for EpCAM, but negative for EMA and Her2/neu. The dashed arrow indicates tumor cells that are positive for Her2/neu, but negative for EpCAM and EMA. Scale bars represent 100 μm. EpCAM, epithelial cell adhesion molecule; EMA, epithelial membrane antigen; Her2/neu, human epidermal growth receptor type 2
    Antibodies Against Her2 Erbb2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Santa Cruz Biotechnology antibodies against her2 neu
    Detection of ovarian metastases by a combination of markers. Representative image of a lobular ovarian metastasis stained with DAPI counterstain and triple immunofluorescence for EpCAM ( a ), EMA ( b ), <t>Her2/neu</t> ( c ), and the three stainings combined ( d ). The solid arrow indicates tumor cells that are positive for EpCAM, but negative for EMA and Her2/neu. The dashed arrow indicates tumor cells that are positive for Her2/neu, but negative for EpCAM and EMA. Scale bars represent 100 μm. EpCAM, epithelial cell adhesion molecule; EMA, epithelial membrane antigen; Her2/neu, human epidermal growth receptor type 2
    Antibodies Against Her2 Neu, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Genentech antibody against her2
    SDS-PAGE of <t>HER2</t> from SK-BR3 cell lysate . Lane 1: Marker; Lane 2: 10 6 SK-BR3 cell lysate; Lane 3: whole protein extract using Norgen kit; Lane 4: control- carboxyl functionalized IO for capture; Lane 5: IO-Ab for capturefrom non-purified SK-BR3 cell
    Antibody Against Her2, supplied by Genentech, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore monoclonal antibody against her2
    Pharmacological blockade of FASN activity suppresses the <t>HER2-driven</t> tumor-initiating capacity of CSCs Mammosphere formation of MCF-7 and MCF-7/HER2 cells with/without C75 (left) or (-)-C75 (right). MSFE of vehicle-alone control cells was normalized to one in each cell line.
    Monoclonal Antibody Against Her2, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Agilent technologies polyclonal antibody against her2 oncoprotein
    Pharmacological blockade of FASN activity suppresses the <t>HER2-driven</t> tumor-initiating capacity of CSCs Mammosphere formation of MCF-7 and MCF-7/HER2 cells with/without C75 (left) or (-)-C75 (right). MSFE of vehicle-alone control cells was normalized to one in each cell line.
    Polyclonal Antibody Against Her2 Oncoprotein, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies polyclonal antibody against her2 protein
    Pharmacological blockade of FASN activity suppresses the <t>HER2-driven</t> tumor-initiating capacity of CSCs Mammosphere formation of MCF-7 and MCF-7/HER2 cells with/without C75 (left) or (-)-C75 (right). MSFE of vehicle-alone control cells was normalized to one in each cell line.
    Polyclonal Antibody Against Her2 Protein, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 90/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Agilent technologies rabbit polyclonal antibody against human her2
    Pharmacological blockade of FASN activity suppresses the <t>HER2-driven</t> tumor-initiating capacity of CSCs Mammosphere formation of MCF-7 and MCF-7/HER2 cells with/without C75 (left) or (-)-C75 (right). MSFE of vehicle-alone control cells was normalized to one in each cell line.
    Rabbit Polyclonal Antibody Against Human Her2, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Agilent technologies monoclonal antibodies against her 2
    Pharmacological blockade of FASN activity suppresses the <t>HER2-driven</t> tumor-initiating capacity of CSCs Mammosphere formation of MCF-7 and MCF-7/HER2 cells with/without C75 (left) or (-)-C75 (right). MSFE of vehicle-alone control cells was normalized to one in each cell line.
    Monoclonal Antibodies Against Her 2, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cell Signaling Technology Inc monoclonal antibodies against her2
    Sulforaphane inhibits multiple cancer development-associated signaling pathways in A2780 cells. (A) Effect of sulforaphane on intracellular levels of <t>Her2</t> through immunofluorescence assays (magnification, ×100). (B) Western blot assays were performed to examine the effect of sulforaphane on the 10 cancer-associated signaling pathway reporter activities, including Bcl-2, Bax, Cyto-c, Caspase-3, p-AKT, p-NF-κB, P53, P27, cyclin-D1 and cMyc. The values are presented as the mean ± standard deviation (n=8-10) of the samples. + P
    Monoclonal Antibodies Against Her2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Ventana Medical primary antibodies against her2
    The HSF1 transcriptional program is impaired by <t>HER2</t> inhibition. ( a ) Schematic overview of HSF1-regulated gene expression. Transcriptionally activated and -repressed target genes analyzed here are indicated. ( b and c ) HER2 inhibition impairs the HSF1-dependent inducible heat-shock response ( b ) and the tumor-promoting ( c ) transcriptional program of HSF1. SK-BR-3 cells were treated with 2 μ M CP724.714 or DMSO for 48 h and mRNA was isolated. qRT-PCR of Hsp70 (* P =0.0396), Hsp110 (*** P =0.001), Hsp90 α (** P =0.0049), SPTAN (* P =0.0362), CDC6 (*** P =0.001), FASN (** P =0.0069) and CBX3 ( P =0.067), each normalized to 36B4 mRNA. Relative values are given in (ratio (2 −ddCT )). Error bars indicate ±S.E.M. of three independent experiments, repeated twice each with all in triplicates. Student's t -test, two-tailed, P- value: * P
    Primary Antibodies Against Her2, supplied by Ventana Medical, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Agilent technologies antibodies against her 2 neu
    The HSF1 transcriptional program is impaired by <t>HER2</t> inhibition. ( a ) Schematic overview of HSF1-regulated gene expression. Transcriptionally activated and -repressed target genes analyzed here are indicated. ( b and c ) HER2 inhibition impairs the HSF1-dependent inducible heat-shock response ( b ) and the tumor-promoting ( c ) transcriptional program of HSF1. SK-BR-3 cells were treated with 2 μ M CP724.714 or DMSO for 48 h and mRNA was isolated. qRT-PCR of Hsp70 (* P =0.0396), Hsp110 (*** P =0.001), Hsp90 α (** P =0.0049), SPTAN (* P =0.0362), CDC6 (*** P =0.001), FASN (** P =0.0069) and CBX3 ( P =0.067), each normalized to 36B4 mRNA. Relative values are given in (ratio (2 −ddCT )). Error bars indicate ±S.E.M. of three independent experiments, repeated twice each with all in triplicates. Student's t -test, two-tailed, P- value: * P
    Antibodies Against Her 2 Neu, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Novocastra antibodies against her 2
    Comparison of gene amplification patterns of <t>Her-2,</t> EGFR and cyclin D1 in primary tumor and the corresponding metastatic lymph node (LN) (×400). High-level amplification/high-level amplification of Her-2 in primary tumor ( A ) and LN ( B ); high-level amplification/no amplification of Her-2 in primary tumor ( C ) and LN ( D ); high-level amplification/no amplification of EGFR in primary tumor ( E ) and LN ( F ); low-level amplification/low-level amplification of EGFR in primary tumor ( G ) and LN ( H ); high-level amplification/no amplification of cyclin D1 in primary tumor ( I ) and LN ( J ); high-level amplification/high-level amplification of cyclin D1 in primary tumor ( K ) and LN ( L ).
    Antibodies Against Her 2, supplied by Novocastra, 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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    Upstate Biotechnology Inc antibodies against her 2
    Representative pictures of <t>HER-2</t> IHC and FISH staining on models EC004, 016, 039, 044 and 054 with matched human primary tumor tissues. HER-2 moderate staining (++) and FISH GCN increase were detected in models EC039 and EC044 and matched primary tumor tissues. EC004, EC016 and EC054 were shown to be HER-2 IHC negative and FISH non-AMP.
    Antibodies Against Her 2, supplied by Upstate Biotechnology Inc, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher antibodies against her2 ecd
    Microscopic analysis of the <t>HER2-ECD-induced</t> HER2-negative cells treated with Tra-IR700-mediated PIT. A, morphological changes of the tumor cells after PIT. MKN1 and MKN45 were cultured with Ad/HER2-ECD for 48 h and were treated with Tra-IR700 for 6 h.
    Antibodies Against Her2 Ecd, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Santa Cruz Biotechnology antibodies against her2 erbb2
    Microscopic analysis of the <t>HER2-ECD-induced</t> HER2-negative cells treated with Tra-IR700-mediated PIT. A, morphological changes of the tumor cells after PIT. MKN1 and MKN45 were cultured with Ad/HER2-ECD for 48 h and were treated with Tra-IR700 for 6 h.
    Antibodies Against Her2 Erbb2, supplied by Santa Cruz Biotechnology, 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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    Image Search Results


    Western blots confirmed low (MCF7 parental) intermediate (Clone A) and high (Clone B) Her2 expression. On the left (no 17-DMAG, dose 0), samples of 3 experiments were loaded into 1 gel to show the standard error of the mean. Her2 expression decreased

    Journal: Clinical Cancer Research

    Article Title: Optical Imaging with HER2-targeted Affibody Molecules can monitor Hsp90 treatment response in a breast cancer xenograft mouse model

    doi: 10.1158/1078-0432.CCR-10-3213

    Figure Lengend Snippet: Western blots confirmed low (MCF7 parental) intermediate (Clone A) and high (Clone B) Her2 expression. On the left (no 17-DMAG, dose 0), samples of 3 experiments were loaded into 1 gel to show the standard error of the mean. Her2 expression decreased

    Article Snippet: After incubation with primary antibodies against Her2 (c-erbB-2 Ab-17, Thermo Fisher Scientific, Fremont, CA) and α-Tubulin (clone B-5-1-2, Sigma-Aldrich Inc. ), and development of the blot, the protein density bands were analyzed using ImageJ software (Version 1.41, National Institutes of Health, Bethesda, MD).

    Techniques: Western Blot, Expressing

    Flow cytometry results showed low (MCF7 parental; ———), intermediate (Clone A; ---), and high (Clone B; ·····) expression levels of Her2, respectively, in comparison with the control MCF7

    Journal: Clinical Cancer Research

    Article Title: Optical Imaging with HER2-targeted Affibody Molecules can monitor Hsp90 treatment response in a breast cancer xenograft mouse model

    doi: 10.1158/1078-0432.CCR-10-3213

    Figure Lengend Snippet: Flow cytometry results showed low (MCF7 parental; ———), intermediate (Clone A; ---), and high (Clone B; ·····) expression levels of Her2, respectively, in comparison with the control MCF7

    Article Snippet: After incubation with primary antibodies against Her2 (c-erbB-2 Ab-17, Thermo Fisher Scientific, Fremont, CA) and α-Tubulin (clone B-5-1-2, Sigma-Aldrich Inc. ), and development of the blot, the protein density bands were analyzed using ImageJ software (Version 1.41, National Institutes of Health, Bethesda, MD).

    Techniques: Flow Cytometry, Cytometry, Expressing

    Figure 4A: Western blotting analysis of LTLT-Ca cells treated with entinostat alone or in presence of MG-132 (proteosomal inhibitor) or NH 4 Cl (lysosomal inhibitor): LTLT-Ca cells were treated with ENT (1μM) alone or in presence of MG-132 (5μM) or NH 4 Cl (100μM) or both. Protein expression in the cells was examined by western immunoblotting as described in “materials and methods”. Blots were probed for β-actin to verify equal loading. The numbers below the blots show densitometric values that are corrected for loading. Figure 4B: Western blotting analysis of SKBr3 and BT-474 cells treated with entinostat: Her-2 positive SKBr3 and BT-474 cells were treated with ENT (1μM). Protein expression in the cells was examined by western immunoblotting as described in “materials and methods”. Blots were probed for β-actin to verify equal loading. The numbers below the blots show densitometric values that are corrected for loading. Figure 4C: Western blotting analysis of AnR and ExR cells treated with entinostat: Her-2 positive anastrozole resistant (AnR) and exemestane resistant (ExR) cells were treated with ENT (1μM). Protein expression in the cells was examined by western immunoblotting as described in “materials and methods”. Blots were probed for β-actin to verify equal loading. The numbers below the blots show densitometric values that are corrected for loading.

    Journal: Molecular cancer therapeutics

    Article Title: HDAC inhibitor entinostat restores responsiveness of letrozole resistant MCF-7Ca xenografts to AIs through modulation of Her-2

    doi: 10.1158/1535-7163.MCT-13-0345

    Figure Lengend Snippet: Figure 4A: Western blotting analysis of LTLT-Ca cells treated with entinostat alone or in presence of MG-132 (proteosomal inhibitor) or NH 4 Cl (lysosomal inhibitor): LTLT-Ca cells were treated with ENT (1μM) alone or in presence of MG-132 (5μM) or NH 4 Cl (100μM) or both. Protein expression in the cells was examined by western immunoblotting as described in “materials and methods”. Blots were probed for β-actin to verify equal loading. The numbers below the blots show densitometric values that are corrected for loading. Figure 4B: Western blotting analysis of SKBr3 and BT-474 cells treated with entinostat: Her-2 positive SKBr3 and BT-474 cells were treated with ENT (1μM). Protein expression in the cells was examined by western immunoblotting as described in “materials and methods”. Blots were probed for β-actin to verify equal loading. The numbers below the blots show densitometric values that are corrected for loading. Figure 4C: Western blotting analysis of AnR and ExR cells treated with entinostat: Her-2 positive anastrozole resistant (AnR) and exemestane resistant (ExR) cells were treated with ENT (1μM). Protein expression in the cells was examined by western immunoblotting as described in “materials and methods”. Blots were probed for β-actin to verify equal loading. The numbers below the blots show densitometric values that are corrected for loading.

    Article Snippet: Antibodies against Her-2 and p-Her-2 were purchased from Millipore (Billerica, MA); antibodies against p-MAPK, MAPK, p-Elk-1, and p-p90RSK were purchased from Cell Signaling Technology (Beverly, MA).

    Techniques: Western Blot, Expressing

    Figure 6A: RT-PCR analysis of Her-2 mRNA levels in LTLT-Ca cells: Relative levels of Her-2 mRNA after treatment with ENT were analyzed by real-time RT-qPCR. Cells were treated with ENT (1μM) alone or in presence of actinomycin D (5μM) and cells were collected at indicated time points. RNA was isolated, quantified and diluted to 0.08μg/μl. Total 0.64Mg of RNA was reverse transcribed and Her-2 was amplified using real-time qPCR as described in Materials and Methods. Graph represents relative levels (±SEM) of Her-2 mRNA compared to LTLT-Ca control (set at 1). Figure 6B, C: RT-PCR analysis of Her-2, HDAC1, ERα and 18s mRNA levels in LTLT-Ca cells: Relative levels of Her-2, HDAC1, ERα and 18s mRNA after treatment with ENT, or siRNA against HDAC1 were analyzed by real-time RT-qPCR. Cells were treated with ENT (1μM) or siRNAs against HDAC1 (5nM) or mock siRNA. The cells were collected after 48-hour incubation at 37°C. RNA was isolated, quantified and diluted to 0.08μg/μl. Total 0.64Mg of RNA was reverse transcribed and cDNA was amplified using real-time qPCR as described in Materials and Methods. Graph represents relative levels (±SEM) of (B) Her-2 and HDAC1 mRNA and (C) ERα and 18s mRNA. The relative mRNA levels (±SEM) are reported compared to LTLT-Ca control (set at 1). P-values: *p=0.0024, ‡p=0.0021, †p

    Journal: Molecular cancer therapeutics

    Article Title: HDAC inhibitor entinostat restores responsiveness of letrozole resistant MCF-7Ca xenografts to AIs through modulation of Her-2

    doi: 10.1158/1535-7163.MCT-13-0345

    Figure Lengend Snippet: Figure 6A: RT-PCR analysis of Her-2 mRNA levels in LTLT-Ca cells: Relative levels of Her-2 mRNA after treatment with ENT were analyzed by real-time RT-qPCR. Cells were treated with ENT (1μM) alone or in presence of actinomycin D (5μM) and cells were collected at indicated time points. RNA was isolated, quantified and diluted to 0.08μg/μl. Total 0.64Mg of RNA was reverse transcribed and Her-2 was amplified using real-time qPCR as described in Materials and Methods. Graph represents relative levels (±SEM) of Her-2 mRNA compared to LTLT-Ca control (set at 1). Figure 6B, C: RT-PCR analysis of Her-2, HDAC1, ERα and 18s mRNA levels in LTLT-Ca cells: Relative levels of Her-2, HDAC1, ERα and 18s mRNA after treatment with ENT, or siRNA against HDAC1 were analyzed by real-time RT-qPCR. Cells were treated with ENT (1μM) or siRNAs against HDAC1 (5nM) or mock siRNA. The cells were collected after 48-hour incubation at 37°C. RNA was isolated, quantified and diluted to 0.08μg/μl. Total 0.64Mg of RNA was reverse transcribed and cDNA was amplified using real-time qPCR as described in Materials and Methods. Graph represents relative levels (±SEM) of (B) Her-2 and HDAC1 mRNA and (C) ERα and 18s mRNA. The relative mRNA levels (±SEM) are reported compared to LTLT-Ca control (set at 1). P-values: *p=0.0024, ‡p=0.0021, †p

    Article Snippet: Antibodies against Her-2 and p-Her-2 were purchased from Millipore (Billerica, MA); antibodies against p-MAPK, MAPK, p-Elk-1, and p-p90RSK were purchased from Cell Signaling Technology (Beverly, MA).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Isolation, Amplification, Real-time Polymerase Chain Reaction, Incubation

    Let-7f regulates β2-AR expression in breast cancer cells. a to d , MCF-7 ( a and b ) and MCF-7/Her2 cells ( c and d ) were planted in 24-well plates and transfected with 9 and 27 pmol synthetic inhibitors or mimics of let-7f. The expression of β2-AR was analyzed by Western blot. These experiments were repeated twice

    Journal: BMC Cancer

    Article Title: A Her2-let-7-β2-AR circuit affects prognosis in patients with Her2-positive breast cancer

    doi: 10.1186/s12885-015-1869-6

    Figure Lengend Snippet: Let-7f regulates β2-AR expression in breast cancer cells. a to d , MCF-7 ( a and b ) and MCF-7/Her2 cells ( c and d ) were planted in 24-well plates and transfected with 9 and 27 pmol synthetic inhibitors or mimics of let-7f. The expression of β2-AR was analyzed by Western blot. These experiments were repeated twice

    Article Snippet: The antibodies were used for immunoblotting: the antibodies against Her2 (4290, Cell Signaling), p-ERK (4370, Cell Signaling), ERK (4695, Cell Signaling), β2-AR (sc-569, Santa Cruiz), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Sungene Biotech).

    Techniques: Expressing, Transfection, Western Blot

    β2-AR overexpression correlates with DFS and LNM in breast cancer patients. a and b , The rates of DFS ( a ) and OS ( b ) in the patients with Her2-positive metastatic breast cancer according to the expression level of β2-AR were determined by the Kaplan-Meier analysis. c , The expression of Her2 and β2-AR was analyzed using a tissue microarray containing 50 metastatic lymph nodes from breast cancer patients by immunohistochemical staining. The middle and right panels are the magnifications of the square regions in the left and middle panels, respectively. Bar = 1000 μm (low-power field), 200 μm or 100 μm (high-power field). d , The relationship between LNM and β2-AR expression was evaluated in Her2-overexpressing breast cancer

    Journal: BMC Cancer

    Article Title: A Her2-let-7-β2-AR circuit affects prognosis in patients with Her2-positive breast cancer

    doi: 10.1186/s12885-015-1869-6

    Figure Lengend Snippet: β2-AR overexpression correlates with DFS and LNM in breast cancer patients. a and b , The rates of DFS ( a ) and OS ( b ) in the patients with Her2-positive metastatic breast cancer according to the expression level of β2-AR were determined by the Kaplan-Meier analysis. c , The expression of Her2 and β2-AR was analyzed using a tissue microarray containing 50 metastatic lymph nodes from breast cancer patients by immunohistochemical staining. The middle and right panels are the magnifications of the square regions in the left and middle panels, respectively. Bar = 1000 μm (low-power field), 200 μm or 100 μm (high-power field). d , The relationship between LNM and β2-AR expression was evaluated in Her2-overexpressing breast cancer

    Article Snippet: The antibodies were used for immunoblotting: the antibodies against Her2 (4290, Cell Signaling), p-ERK (4370, Cell Signaling), ERK (4695, Cell Signaling), β2-AR (sc-569, Santa Cruiz), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Sungene Biotech).

    Techniques: Over Expression, Expressing, Microarray, Immunohistochemistry, Staining

    Her2 overexpression inhibits let-7f via constitutive activation of ERK. a , The expression of let-7f in MCF-7 and MCF-7/Her2 cells was detected by real-time RT-PCR. b and c , The expression of Her2, β2-AR, and phosphorylated ERK in MCF-7 and SKBR3 cells was analyzed by Western blot ( b ) and the level of let-7f was detected by real-time RT-PCR ( c ). d and e , SKBR3 cells were transfected with the siRNA targeting Her2. The expression of Her2, β2-AR, and phosphorylated ERK was analyzed by Western blot ( d ) and the level of let-7f was detected by real-time RT-PCR ( e ). f , MCF-7/Her2 cells were pre-treated with 25 μM PD98059 or DMSO (as a solvent control) for 24 h and the expression of let-7f was analyzed. g and h , MCF-7/Her2 cells were pre-treated with 1 μM PD184352 ( g ) or 0.5 μM GDC0941 (h) for 2 h. The levels of phosphorylated ERK and AKT were analyzed by Western blot and the expression of let-7f was detected by real-time RT-PCR. i , MCF-7/Her2 cells were treated with 2.5 μM ISO and the expression of let-7f was analyzed by real-time RT-PCR. These experiments were repeated at least twice. * P

    Journal: BMC Cancer

    Article Title: A Her2-let-7-β2-AR circuit affects prognosis in patients with Her2-positive breast cancer

    doi: 10.1186/s12885-015-1869-6

    Figure Lengend Snippet: Her2 overexpression inhibits let-7f via constitutive activation of ERK. a , The expression of let-7f in MCF-7 and MCF-7/Her2 cells was detected by real-time RT-PCR. b and c , The expression of Her2, β2-AR, and phosphorylated ERK in MCF-7 and SKBR3 cells was analyzed by Western blot ( b ) and the level of let-7f was detected by real-time RT-PCR ( c ). d and e , SKBR3 cells were transfected with the siRNA targeting Her2. The expression of Her2, β2-AR, and phosphorylated ERK was analyzed by Western blot ( d ) and the level of let-7f was detected by real-time RT-PCR ( e ). f , MCF-7/Her2 cells were pre-treated with 25 μM PD98059 or DMSO (as a solvent control) for 24 h and the expression of let-7f was analyzed. g and h , MCF-7/Her2 cells were pre-treated with 1 μM PD184352 ( g ) or 0.5 μM GDC0941 (h) for 2 h. The levels of phosphorylated ERK and AKT were analyzed by Western blot and the expression of let-7f was detected by real-time RT-PCR. i , MCF-7/Her2 cells were treated with 2.5 μM ISO and the expression of let-7f was analyzed by real-time RT-PCR. These experiments were repeated at least twice. * P

    Article Snippet: The antibodies were used for immunoblotting: the antibodies against Her2 (4290, Cell Signaling), p-ERK (4370, Cell Signaling), ERK (4695, Cell Signaling), β2-AR (sc-569, Santa Cruiz), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Sungene Biotech).

    Techniques: Over Expression, Activation Assay, Expressing, Quantitative RT-PCR, Western Blot, Transfection

    β2-AR is highly expressed in Her2-positive breast cancer tissues. a and b , The relative mRNA expression of Her2 ( a ) and ADRB2 ( b ) in human breast cancer (2, n = 53) and normal breast tissue samples (1, n = 6) was analyzed by searching a publicly available database Oncomine ( www.oncomine.org ). c , The expression of Her2 and β2-AR was detected by immunohistochemistry on a human breast cancer tissue microarray consisting of 49 tumor tissues from breast cancer patients. Bar = 1000 μm (low-power field) or 100 μm (high-power field)

    Journal: BMC Cancer

    Article Title: A Her2-let-7-β2-AR circuit affects prognosis in patients with Her2-positive breast cancer

    doi: 10.1186/s12885-015-1869-6

    Figure Lengend Snippet: β2-AR is highly expressed in Her2-positive breast cancer tissues. a and b , The relative mRNA expression of Her2 ( a ) and ADRB2 ( b ) in human breast cancer (2, n = 53) and normal breast tissue samples (1, n = 6) was analyzed by searching a publicly available database Oncomine ( www.oncomine.org ). c , The expression of Her2 and β2-AR was detected by immunohistochemistry on a human breast cancer tissue microarray consisting of 49 tumor tissues from breast cancer patients. Bar = 1000 μm (low-power field) or 100 μm (high-power field)

    Article Snippet: The antibodies were used for immunoblotting: the antibodies against Her2 (4290, Cell Signaling), p-ERK (4370, Cell Signaling), ERK (4695, Cell Signaling), β2-AR (sc-569, Santa Cruiz), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Sungene Biotech).

    Techniques: Expressing, Immunohistochemistry, Microarray

    Immunohistochemistry of Her2 and GEP100 in adenocarcinoma lung tissue specimens. ( A ) Her2 and GEP100 were each stained in brown in sequential sections. Case 1, Her2 and GEP100 double negative; Case 2, Her2 score 1+ and GEP100 score 1+; Case 3, Her2 score 2+ and GEP100 score 2+; Case 4, Her2 score 3+ GEP100 and GEP100 score 2+. All sections were counterstained with hematoxylin and eosin (H E) and representative figures are shown. Scale bars, 100 µm. ( B ) Representative figures of the peripheral expression of GEP100. Scale bars, 500 µm. ( C ) No correlation between strong expression of GEP100 (score 2+) per se with node-metastases. ( D ) A statistical correlation between double strong positive signal of GEP100 (score 2+) and Her2 (score 3+), and node-metastases. In C and D , solid bars mean GEP100 positive rate for strong expression (score 2+).

    Journal: PLoS ONE

    Article Title: Engagement of Overexpressed Her2 with GEP100 Induces Autonomous Invasive Activities and Provides a Biomarker for Metastases of Lung Adenocarcinoma

    doi: 10.1371/journal.pone.0025301

    Figure Lengend Snippet: Immunohistochemistry of Her2 and GEP100 in adenocarcinoma lung tissue specimens. ( A ) Her2 and GEP100 were each stained in brown in sequential sections. Case 1, Her2 and GEP100 double negative; Case 2, Her2 score 1+ and GEP100 score 1+; Case 3, Her2 score 2+ and GEP100 score 2+; Case 4, Her2 score 3+ GEP100 and GEP100 score 2+. All sections were counterstained with hematoxylin and eosin (H E) and representative figures are shown. Scale bars, 100 µm. ( B ) Representative figures of the peripheral expression of GEP100. Scale bars, 500 µm. ( C ) No correlation between strong expression of GEP100 (score 2+) per se with node-metastases. ( D ) A statistical correlation between double strong positive signal of GEP100 (score 2+) and Her2 (score 3+), and node-metastases. In C and D , solid bars mean GEP100 positive rate for strong expression (score 2+).

    Article Snippet: We used the same antibody against Her2 as used in the Herceptest (DAKO, A0485).

    Techniques: Immunohistochemistry, Staining, Expressing

    GEP100 associates with Her2 to induce Arf6 activation. ( A ) Co-precipitation of Her2-EGFP with HA-GEP100, expressed in 293T cells, and analysed by anti-GEP100 immunoprecipitation (IP) coupled with anti-GFP immunoblots (IB). Anti-GEP100 immunoprecipitants were also blotted by anti-phospho Her2 and an anti-HA antibody. Immunoprecipitation for the expression of Her2-EGFP without HA-GEP100 was included as a control (vector). Immunoprecipitation using non-immune serum was also included as a control (NC). ( B ) In vitro co-precipitation of Her2-EGFP (pEGFP-Her2) expressed in cells with the two indicated GST-tagged PH domains (GST-GEP100-PH/GST-ARNO-PH) or GST alone, analysed by glutathione-beads pulldown and anti-GFP immunoblots. GST-fusion proteins were visualized by Ponceau S. In A and B , cells were cultured with 10% FCS (Se), in the absence of serum for 24 h (St), or stimulated with 10 ng/ml EGF for 10 min after serum starvation for 24 h (E), prior to lysis. EGFP alone was included as a control (pEGFP). ( C ) Co-precipitation of HA-GEP100 with wild type Her2-EGFP (WT) or its mutants (YF1; 1139F, YF2; 1196F, YF3; 1221/1222F, YF4; 1248F, YF1/2; 1139/1196F, YF3/4; 1221/1222/1248F), analysed by anti-GEP100 immunoprecipitation and anti-GFP immunoblots. ( D ) Arf6-myc activities in cells expressing HA-GEP100 and Her2-EGFP or their mutants, measured by GST-GGA pulldown and anti-myc immunoblots. +, wild type; YF, Her2 1139/1196F mutant; Del, Sec7-deleted GEP100. EGFP alone was included as a control (G). In A – D , immunoblots of total cell lysates (10 µg) are also shown (Total).

    Journal: PLoS ONE

    Article Title: Engagement of Overexpressed Her2 with GEP100 Induces Autonomous Invasive Activities and Provides a Biomarker for Metastases of Lung Adenocarcinoma

    doi: 10.1371/journal.pone.0025301

    Figure Lengend Snippet: GEP100 associates with Her2 to induce Arf6 activation. ( A ) Co-precipitation of Her2-EGFP with HA-GEP100, expressed in 293T cells, and analysed by anti-GEP100 immunoprecipitation (IP) coupled with anti-GFP immunoblots (IB). Anti-GEP100 immunoprecipitants were also blotted by anti-phospho Her2 and an anti-HA antibody. Immunoprecipitation for the expression of Her2-EGFP without HA-GEP100 was included as a control (vector). Immunoprecipitation using non-immune serum was also included as a control (NC). ( B ) In vitro co-precipitation of Her2-EGFP (pEGFP-Her2) expressed in cells with the two indicated GST-tagged PH domains (GST-GEP100-PH/GST-ARNO-PH) or GST alone, analysed by glutathione-beads pulldown and anti-GFP immunoblots. GST-fusion proteins were visualized by Ponceau S. In A and B , cells were cultured with 10% FCS (Se), in the absence of serum for 24 h (St), or stimulated with 10 ng/ml EGF for 10 min after serum starvation for 24 h (E), prior to lysis. EGFP alone was included as a control (pEGFP). ( C ) Co-precipitation of HA-GEP100 with wild type Her2-EGFP (WT) or its mutants (YF1; 1139F, YF2; 1196F, YF3; 1221/1222F, YF4; 1248F, YF1/2; 1139/1196F, YF3/4; 1221/1222/1248F), analysed by anti-GEP100 immunoprecipitation and anti-GFP immunoblots. ( D ) Arf6-myc activities in cells expressing HA-GEP100 and Her2-EGFP or their mutants, measured by GST-GGA pulldown and anti-myc immunoblots. +, wild type; YF, Her2 1139/1196F mutant; Del, Sec7-deleted GEP100. EGFP alone was included as a control (G). In A – D , immunoblots of total cell lysates (10 µg) are also shown (Total).

    Article Snippet: We used the same antibody against Her2 as used in the Herceptest (DAKO, A0485).

    Techniques: Activation Assay, Immunoprecipitation, Western Blot, Expressing, Plasmid Preparation, In Vitro, Cell Culture, Lysis, Mutagenesis

    Endogenous binding of GEP100 with Her2 overexpressed in H522 lung adenocarcinoma cells. ( A ) Total lysates (30 µg) of 11 non-small cell lung cancer cell lines were immunoblotted with antibodies as indicated. ( B ) Co-precipitation of Her2 with GEP100 from H522 cell lysates (250 µg), analysed by anti-GEP100 immunoprecipitation, and the immunoblot for anti-GEP100, anti-Her2 (HER2), and anti-phospho Her2 (pHER2). Immunoblots using anti-EGFR (EGFR) and anti-Her3 (HER3) antibodies were also included. Cells were cultured in the presence of serum (Se), or starved for serum for 24 h (St), prior to lysis. Total, 20 µg of total lysates. Immunoprecipitation of cells using non-immune serum was used as a negative control (NC).

    Journal: PLoS ONE

    Article Title: Engagement of Overexpressed Her2 with GEP100 Induces Autonomous Invasive Activities and Provides a Biomarker for Metastases of Lung Adenocarcinoma

    doi: 10.1371/journal.pone.0025301

    Figure Lengend Snippet: Endogenous binding of GEP100 with Her2 overexpressed in H522 lung adenocarcinoma cells. ( A ) Total lysates (30 µg) of 11 non-small cell lung cancer cell lines were immunoblotted with antibodies as indicated. ( B ) Co-precipitation of Her2 with GEP100 from H522 cell lysates (250 µg), analysed by anti-GEP100 immunoprecipitation, and the immunoblot for anti-GEP100, anti-Her2 (HER2), and anti-phospho Her2 (pHER2). Immunoblots using anti-EGFR (EGFR) and anti-Her3 (HER3) antibodies were also included. Cells were cultured in the presence of serum (Se), or starved for serum for 24 h (St), prior to lysis. Total, 20 µg of total lysates. Immunoprecipitation of cells using non-immune serum was used as a negative control (NC).

    Article Snippet: We used the same antibody against Her2 as used in the Herceptest (DAKO, A0485).

    Techniques: Binding Assay, Immunoprecipitation, Western Blot, Cell Culture, Lysis, Negative Control

    Inhibition of the Her2-GEP100 pathway blocks invasive activities of H522 cells. ( A ) Cells, untreated or treated with AG825 (10 µM) or DMSO (1%) for 72 h, were subjected to the Matrigel invasion assay, or their lysates (30 µg) were analysed by immunoblotting as indicated. Cell viability, measured by the MTS assay, is also shown. ( B ) Cells, untransfected (mock), or transfected with siRNA duplexes against GEP100 or irrelevant sequences (Irr), were subjected to the Matrigel invasion assay and the MTS assay; or were analysed for their expression of GEP100 by immunoblotting of the lysates (20 µg) using the antibody as indicated. β-actin immunoblots were included as controls. ( C ) Cells, expressing EGFP-tagged GEP100 PH domain (EGFP-GEP100-PH) or EGFP alone (EGFP), were subjected to the Matrigel invasion assay and the MTS assay. Inhibition of the intracellular association of GEP100 with Her2 by EGFP-GEP100-PH, as well as the association of EGFP-GEP100-PH with endogenous Her2, were confirmed by anti-GEP100 and anti-GFP immunoprecipitation coupled with anti-Her2 immunoblot, as indicated. IP, immunoprecipitant for total cell lysates (250 µg). Total, total cell lysates (30 µg). ( D ) Cells, untransfected (mock) or transfected with siRNA duplexes against Arf6, AMAP1, or with irrelevant sequences (Irr), were subjected to the Matrigel invasion assay and the MTS assay; or were analysed for their expression of Arf6 and AMAP1 by immunoblotting of the lysates using the antibodies as indicated. β-actin immunoblots were included as controls. In each bar chart, data of invasive activities (black columns) and cell viabilities (white columns) are presented as percentages calculated by normalizing the values obtained for the untreated cells (3A) which means native cells, or the mock cells (3B, 3C, and 3D) to which only transfection reagents were added as 1.0. Error bars show mean +/− s.e.m., n = 3. *, p

    Journal: PLoS ONE

    Article Title: Engagement of Overexpressed Her2 with GEP100 Induces Autonomous Invasive Activities and Provides a Biomarker for Metastases of Lung Adenocarcinoma

    doi: 10.1371/journal.pone.0025301

    Figure Lengend Snippet: Inhibition of the Her2-GEP100 pathway blocks invasive activities of H522 cells. ( A ) Cells, untreated or treated with AG825 (10 µM) or DMSO (1%) for 72 h, were subjected to the Matrigel invasion assay, or their lysates (30 µg) were analysed by immunoblotting as indicated. Cell viability, measured by the MTS assay, is also shown. ( B ) Cells, untransfected (mock), or transfected with siRNA duplexes against GEP100 or irrelevant sequences (Irr), were subjected to the Matrigel invasion assay and the MTS assay; or were analysed for their expression of GEP100 by immunoblotting of the lysates (20 µg) using the antibody as indicated. β-actin immunoblots were included as controls. ( C ) Cells, expressing EGFP-tagged GEP100 PH domain (EGFP-GEP100-PH) or EGFP alone (EGFP), were subjected to the Matrigel invasion assay and the MTS assay. Inhibition of the intracellular association of GEP100 with Her2 by EGFP-GEP100-PH, as well as the association of EGFP-GEP100-PH with endogenous Her2, were confirmed by anti-GEP100 and anti-GFP immunoprecipitation coupled with anti-Her2 immunoblot, as indicated. IP, immunoprecipitant for total cell lysates (250 µg). Total, total cell lysates (30 µg). ( D ) Cells, untransfected (mock) or transfected with siRNA duplexes against Arf6, AMAP1, or with irrelevant sequences (Irr), were subjected to the Matrigel invasion assay and the MTS assay; or were analysed for their expression of Arf6 and AMAP1 by immunoblotting of the lysates using the antibodies as indicated. β-actin immunoblots were included as controls. In each bar chart, data of invasive activities (black columns) and cell viabilities (white columns) are presented as percentages calculated by normalizing the values obtained for the untreated cells (3A) which means native cells, or the mock cells (3B, 3C, and 3D) to which only transfection reagents were added as 1.0. Error bars show mean +/− s.e.m., n = 3. *, p

    Article Snippet: We used the same antibody against Her2 as used in the Herceptest (DAKO, A0485).

    Techniques: Inhibition, Invasion Assay, MTS Assay, Transfection, Expressing, Western Blot, Immunoprecipitation

    (A) Expression of HER-2 mRNA, as detected by reverse transcription-quantitative polymerase chain reaction following transfection with the lentiviral-mediated HER2-small hairpin RNA. Differences in the expression of HER-2 mRNA between the KD and NC and CON groups were significant (P

    Journal: Oncology Letters

    Article Title: Suppression of human epidermal growth factor receptor 2 via interference increases the chemosensitivity of ovarian carcinoma

    doi: 10.3892/ol.2016.4341

    Figure Lengend Snippet: (A) Expression of HER-2 mRNA, as detected by reverse transcription-quantitative polymerase chain reaction following transfection with the lentiviral-mediated HER2-small hairpin RNA. Differences in the expression of HER-2 mRNA between the KD and NC and CON groups were significant (P

    Article Snippet: The membranes were then incubated with the primary antibodies against HER2 (rabbit polyclonal; catalog no., ab58616; dilution, 1:150; Abcam, Cambridge, MA, USA) and β-actin (mouse monoclonal; catalog no., AA128; dilution, 1:1,000; Beyotime Institute of Biotechnology, Shanghai, China), followed by incubation with the appropriate goat monoclonal anti-rabbit and rabbit polyclonal anti-mouse secondary antibodies conjugated with alkaline phosphatase (catalog nos., A0208 and A0216, respectively; dilution, 1:1,000; Beyotime Institute of Biotechnology).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection

    (A) Fluorescence expression of SKOV3 cell transfection with the lentiviral-mediated HER2-shRNA. Magnification, ×200. (B) Fluorescent flow cytometry detected fluorescence expression of SKOV3 cell transfection with lentiviral-mediated HER2-shRNA. CON, blank control; NC, negative control; KD, SKOV3 RNA interference cell line; shRNA, small hairpin RNA; FITC, fluorescein isothiocyanate; SS, side scatter; FS, forward scatter; HER2, human epidermal growth factor receptor 2.

    Journal: Oncology Letters

    Article Title: Suppression of human epidermal growth factor receptor 2 via interference increases the chemosensitivity of ovarian carcinoma

    doi: 10.3892/ol.2016.4341

    Figure Lengend Snippet: (A) Fluorescence expression of SKOV3 cell transfection with the lentiviral-mediated HER2-shRNA. Magnification, ×200. (B) Fluorescent flow cytometry detected fluorescence expression of SKOV3 cell transfection with lentiviral-mediated HER2-shRNA. CON, blank control; NC, negative control; KD, SKOV3 RNA interference cell line; shRNA, small hairpin RNA; FITC, fluorescein isothiocyanate; SS, side scatter; FS, forward scatter; HER2, human epidermal growth factor receptor 2.

    Article Snippet: The membranes were then incubated with the primary antibodies against HER2 (rabbit polyclonal; catalog no., ab58616; dilution, 1:150; Abcam, Cambridge, MA, USA) and β-actin (mouse monoclonal; catalog no., AA128; dilution, 1:1,000; Beyotime Institute of Biotechnology, Shanghai, China), followed by incubation with the appropriate goat monoclonal anti-rabbit and rabbit polyclonal anti-mouse secondary antibodies conjugated with alkaline phosphatase (catalog nos., A0208 and A0216, respectively; dilution, 1:1,000; Beyotime Institute of Biotechnology).

    Techniques: Fluorescence, Expressing, Transfection, shRNA, Flow Cytometry, Cytometry, Negative Control

    (A) Significant differences in tumor suppression in mice inoculated with the lentiviral-mediated HER2-shRNA SKOV3 cells and treated with DDP compared with the other four groups (*P

    Journal: Oncology Letters

    Article Title: Suppression of human epidermal growth factor receptor 2 via interference increases the chemosensitivity of ovarian carcinoma

    doi: 10.3892/ol.2016.4341

    Figure Lengend Snippet: (A) Significant differences in tumor suppression in mice inoculated with the lentiviral-mediated HER2-shRNA SKOV3 cells and treated with DDP compared with the other four groups (*P

    Article Snippet: The membranes were then incubated with the primary antibodies against HER2 (rabbit polyclonal; catalog no., ab58616; dilution, 1:150; Abcam, Cambridge, MA, USA) and β-actin (mouse monoclonal; catalog no., AA128; dilution, 1:1,000; Beyotime Institute of Biotechnology, Shanghai, China), followed by incubation with the appropriate goat monoclonal anti-rabbit and rabbit polyclonal anti-mouse secondary antibodies conjugated with alkaline phosphatase (catalog nos., A0208 and A0216, respectively; dilution, 1:1,000; Beyotime Institute of Biotechnology).

    Techniques: Mouse Assay, shRNA

    HBx increased HER2 protein expression by HuR-dependent mRNA stabilization in HCC cells. (a) The HuR protein expression in HBx-paired HCC cells was examined by Western blot ( N = 4). (b) Hep3Bx cells were transiently transfected with either si-control or si-HuR for 4 days. The protein expressions of HER2, EGFR, and HuR were analyzed by Western blot ( N = 3). (c) Hep3Bx cells were transiently transfected with either si-control or si-HuR for 4 days, followed by the treatment of 5 μ M Actinomycin D. The relative remaining HER2 mRNA expression in each group was determined by RT-qPCR. The HER2 mRNA expression was normalized to actin expression. Statistical analysis was performed by Student's t -test. * P

    Journal: BioMed Research International

    Article Title: Hepatitis B Virus X Upregulates HuR Protein Level to Stabilize HER2 Expression in Hepatocellular Carcinoma Cells

    doi: 10.1155/2014/827415

    Figure Lengend Snippet: HBx increased HER2 protein expression by HuR-dependent mRNA stabilization in HCC cells. (a) The HuR protein expression in HBx-paired HCC cells was examined by Western blot ( N = 4). (b) Hep3Bx cells were transiently transfected with either si-control or si-HuR for 4 days. The protein expressions of HER2, EGFR, and HuR were analyzed by Western blot ( N = 3). (c) Hep3Bx cells were transiently transfected with either si-control or si-HuR for 4 days, followed by the treatment of 5 μ M Actinomycin D. The relative remaining HER2 mRNA expression in each group was determined by RT-qPCR. The HER2 mRNA expression was normalized to actin expression. Statistical analysis was performed by Student's t -test. * P

    Article Snippet: We purchased antibodies against HER2, EGFR, and HuR as well as bortezomib from Santa Cruz (Santa Cruz, CA).

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

    The increased HER2 protein expression was responsible for the migration ability of HBx-expressing HCC cells. ((a), (b), (c), and (e)) Hep3Bx cells were transiently transfected with either si-control or si-HER2 for 4 days. Then, cells were either harvested or reseeded for further experiments. Gene silence of HER2 expression was confirmed by Western blot (a) ( N = 3). The relative growth rate was determined by crystal violet staining (b) ( N = 3). The migration of Hep3Bx cells was examined by Transwell migration assay for 48 hrs. The representative pictures of migrated cells were visualized and quantified (c) ( N = 3). The expressions of metastatic factors were examined by Western blot (e) ( N = 3). (d) The expressions of metastatic factors in both Hep3B and Hep3Bx cells were examined by Western blot ( N = 3).

    Journal: BioMed Research International

    Article Title: Hepatitis B Virus X Upregulates HuR Protein Level to Stabilize HER2 Expression in Hepatocellular Carcinoma Cells

    doi: 10.1155/2014/827415

    Figure Lengend Snippet: The increased HER2 protein expression was responsible for the migration ability of HBx-expressing HCC cells. ((a), (b), (c), and (e)) Hep3Bx cells were transiently transfected with either si-control or si-HER2 for 4 days. Then, cells were either harvested or reseeded for further experiments. Gene silence of HER2 expression was confirmed by Western blot (a) ( N = 3). The relative growth rate was determined by crystal violet staining (b) ( N = 3). The migration of Hep3Bx cells was examined by Transwell migration assay for 48 hrs. The representative pictures of migrated cells were visualized and quantified (c) ( N = 3). The expressions of metastatic factors were examined by Western blot (e) ( N = 3). (d) The expressions of metastatic factors in both Hep3B and Hep3Bx cells were examined by Western blot ( N = 3).

    Article Snippet: We purchased antibodies against HER2, EGFR, and HuR as well as bortezomib from Santa Cruz (Santa Cruz, CA).

    Techniques: Expressing, Migration, Transfection, Western Blot, Staining, Transwell Migration Assay

    HBx induced HER2 protein expression in HCC cells. (a) The protein expressions of HER2, HBx, and Tubulin in two HBx-paired HCC cells were examined by Western blot ( N = 4). (b) Myc-HBx expression vector was transiently transfected into Hep3B HCC cells for 48 hrs. The HER2 and myc-HBx protein expressions were analyzed by Western blot ( N = 3). (c) Transient transfection of HBx siRNA was performed in Hep3Bx cells for 4 days. The HER2 protein expression and gene silencing of HBx protein expression were examined by Western blot ( N = 3).

    Journal: BioMed Research International

    Article Title: Hepatitis B Virus X Upregulates HuR Protein Level to Stabilize HER2 Expression in Hepatocellular Carcinoma Cells

    doi: 10.1155/2014/827415

    Figure Lengend Snippet: HBx induced HER2 protein expression in HCC cells. (a) The protein expressions of HER2, HBx, and Tubulin in two HBx-paired HCC cells were examined by Western blot ( N = 4). (b) Myc-HBx expression vector was transiently transfected into Hep3B HCC cells for 48 hrs. The HER2 and myc-HBx protein expressions were analyzed by Western blot ( N = 3). (c) Transient transfection of HBx siRNA was performed in Hep3Bx cells for 4 days. The HER2 protein expression and gene silencing of HBx protein expression were examined by Western blot ( N = 3).

    Article Snippet: We purchased antibodies against HER2, EGFR, and HuR as well as bortezomib from Santa Cruz (Santa Cruz, CA).

    Techniques: Expressing, Western Blot, Plasmid Preparation, Transfection

    The HER2 mRNA expression was stabilized in HBx-expressing HCC cells. (a) The two HBx-paired HCC cells were treated with proteasomal inhibitors (MG132 and bortezomib) for 24 hrs. The HER2 protein expression was analyzed by Western blot ( N = 3). (b) The HER2 mRNA expression in two HBx-paired HCC cells was examined by RT-qPCR. The HER2 mRNA expression was normalized to actin expression. Statistical analysis was performed by Student's t -test. ** P

    Journal: BioMed Research International

    Article Title: Hepatitis B Virus X Upregulates HuR Protein Level to Stabilize HER2 Expression in Hepatocellular Carcinoma Cells

    doi: 10.1155/2014/827415

    Figure Lengend Snippet: The HER2 mRNA expression was stabilized in HBx-expressing HCC cells. (a) The two HBx-paired HCC cells were treated with proteasomal inhibitors (MG132 and bortezomib) for 24 hrs. The HER2 protein expression was analyzed by Western blot ( N = 3). (b) The HER2 mRNA expression in two HBx-paired HCC cells was examined by RT-qPCR. The HER2 mRNA expression was normalized to actin expression. Statistical analysis was performed by Student's t -test. ** P

    Article Snippet: We purchased antibodies against HER2, EGFR, and HuR as well as bortezomib from Santa Cruz (Santa Cruz, CA).

    Techniques: Expressing, Western Blot, Quantitative RT-PCR

    Detection of ovarian metastases by a combination of markers. Representative image of a lobular ovarian metastasis stained with DAPI counterstain and triple immunofluorescence for EpCAM ( a ), EMA ( b ), Her2/neu ( c ), and the three stainings combined ( d ). The solid arrow indicates tumor cells that are positive for EpCAM, but negative for EMA and Her2/neu. The dashed arrow indicates tumor cells that are positive for Her2/neu, but negative for EpCAM and EMA. Scale bars represent 100 μm. EpCAM, epithelial cell adhesion molecule; EMA, epithelial membrane antigen; Her2/neu, human epidermal growth receptor type 2

    Journal: BMC Cancer

    Article Title: Morphological and phenotypical features of ovarian metastases in breast cancer patients

    doi: 10.1186/s12885-017-3191-y

    Figure Lengend Snippet: Detection of ovarian metastases by a combination of markers. Representative image of a lobular ovarian metastasis stained with DAPI counterstain and triple immunofluorescence for EpCAM ( a ), EMA ( b ), Her2/neu ( c ), and the three stainings combined ( d ). The solid arrow indicates tumor cells that are positive for EpCAM, but negative for EMA and Her2/neu. The dashed arrow indicates tumor cells that are positive for Her2/neu, but negative for EpCAM and EMA. Scale bars represent 100 μm. EpCAM, epithelial cell adhesion molecule; EMA, epithelial membrane antigen; Her2/neu, human epidermal growth receptor type 2

    Article Snippet: The sections were incubated overnight in a humidified chamber at room temperature with primary antibodies against Her2/neu (ERBB2, rabbit polyclonal, Dako), E-cadherin (NCH38, mouse monoclonal, Dako), EpCAM (323/A3, mouse monoclonal, provided by the Department of Pathology, LUMC, the Netherlands), CEA (A0115, rabbit polyclonal, Dako), αvβ6 integrin (6.2A1, mouse monoclonal, Cell Essentials), or EMA (E29, mouse monoclonal, Dako); all primary antibodies were used at their predetermined optimal dilution.

    Techniques: Staining, Immunofluorescence

    The correlation between tumor marker expression in breast tumors and ovarian metastases for individual patients. Upper panel ( a ) shows invasive ductal breast cancer and lower panel ( b ) represents invasive lobular breast cancer. For each patient, the percentage of positive tumor cells in primary and locally recurrent breast tumors (if applicable) was set against the percentage of positive tumor cells in their corresponding ovarian metastases. EMA, epithelial membrane antigen; Her2/neu, human epidermal growth receptor type 2; EpCAM, epithelial cell adhesion molecule

    Journal: BMC Cancer

    Article Title: Morphological and phenotypical features of ovarian metastases in breast cancer patients

    doi: 10.1186/s12885-017-3191-y

    Figure Lengend Snippet: The correlation between tumor marker expression in breast tumors and ovarian metastases for individual patients. Upper panel ( a ) shows invasive ductal breast cancer and lower panel ( b ) represents invasive lobular breast cancer. For each patient, the percentage of positive tumor cells in primary and locally recurrent breast tumors (if applicable) was set against the percentage of positive tumor cells in their corresponding ovarian metastases. EMA, epithelial membrane antigen; Her2/neu, human epidermal growth receptor type 2; EpCAM, epithelial cell adhesion molecule

    Article Snippet: The sections were incubated overnight in a humidified chamber at room temperature with primary antibodies against Her2/neu (ERBB2, rabbit polyclonal, Dako), E-cadherin (NCH38, mouse monoclonal, Dako), EpCAM (323/A3, mouse monoclonal, provided by the Department of Pathology, LUMC, the Netherlands), CEA (A0115, rabbit polyclonal, Dako), αvβ6 integrin (6.2A1, mouse monoclonal, Cell Essentials), or EMA (E29, mouse monoclonal, Dako); all primary antibodies were used at their predetermined optimal dilution.

    Techniques: Marker, Expressing

    SDS-PAGE of HER2 from SK-BR3 cell lysate . Lane 1: Marker; Lane 2: 10 6 SK-BR3 cell lysate; Lane 3: whole protein extract using Norgen kit; Lane 4: control- carboxyl functionalized IO for capture; Lane 5: IO-Ab for capturefrom non-purified SK-BR3 cell

    Journal: Biomaterials

    Article Title: Antibody Conjugated Magnetic Iron Oxide Nanoparticles for Cancer Cell Separation in Fresh Whole Blood

    doi: 10.1016/j.biomaterials.2011.08.076

    Figure Lengend Snippet: SDS-PAGE of HER2 from SK-BR3 cell lysate . Lane 1: Marker; Lane 2: 10 6 SK-BR3 cell lysate; Lane 3: whole protein extract using Norgen kit; Lane 4: control- carboxyl functionalized IO for capture; Lane 5: IO-Ab for capturefrom non-purified SK-BR3 cell

    Article Snippet: Antibody against HER2, Anti-HER2, (Herceptin, Genentech, Inc. South San Francisco, CA) was dialyzed in borate buffer and the concentration was established with a Biophotometer (Bio-rad, Philadelphia, PA).

    Techniques: SDS Page, Marker, Purification

    Pharmacological blockade of FASN activity suppresses the HER2-driven tumor-initiating capacity of CSCs Mammosphere formation of MCF-7 and MCF-7/HER2 cells with/without C75 (left) or (-)-C75 (right). MSFE of vehicle-alone control cells was normalized to one in each cell line.

    Journal: Histology and histopathology

    Article Title: Clinical and therapeutic relevance of the metabolic oncogene fatty acid synthase in HER2+ breast cancer

    doi: 10.14670/HH-11-830

    Figure Lengend Snippet: Pharmacological blockade of FASN activity suppresses the HER2-driven tumor-initiating capacity of CSCs Mammosphere formation of MCF-7 and MCF-7/HER2 cells with/without C75 (left) or (-)-C75 (right). MSFE of vehicle-alone control cells was normalized to one in each cell line.

    Article Snippet: HER2 expression was assesed by western blotting using a monoclonal antibody against HER2 (Ab-3 clone, Oncogene Research Products, San Diego, CA) as described ( ).

    Techniques: Activity Assay

    Pharmacological blockade of FASN activity suppresses the HER2-induced transforming phenotype in breast epithelial cells Left. Transcriptional activities were expressed as relative (×-fold) changes in luciferase activity of FASN promoter-transfected MCF10A and MCF10/HER2 cells in response to treatments after normalization to pRL-CMV luciferase activity. Each experimental value represents the mean fold increase ( columns ) ± SD ( bars ) from at least three independent experiments measured in triplicate. Right. Soft-agar colony formation assay of MCF10A and MCF10A/HER2 cells cultured with/without C75. Each experimental value represents the mean colony number ( columns ) ± S. D. ( bars ) from at least three independent experiments measured in triplicate.

    Journal: Histology and histopathology

    Article Title: Clinical and therapeutic relevance of the metabolic oncogene fatty acid synthase in HER2+ breast cancer

    doi: 10.14670/HH-11-830

    Figure Lengend Snippet: Pharmacological blockade of FASN activity suppresses the HER2-induced transforming phenotype in breast epithelial cells Left. Transcriptional activities were expressed as relative (×-fold) changes in luciferase activity of FASN promoter-transfected MCF10A and MCF10/HER2 cells in response to treatments after normalization to pRL-CMV luciferase activity. Each experimental value represents the mean fold increase ( columns ) ± SD ( bars ) from at least three independent experiments measured in triplicate. Right. Soft-agar colony formation assay of MCF10A and MCF10A/HER2 cells cultured with/without C75. Each experimental value represents the mean colony number ( columns ) ± S. D. ( bars ) from at least three independent experiments measured in triplicate.

    Article Snippet: HER2 expression was assesed by western blotting using a monoclonal antibody against HER2 (Ab-3 clone, Oncogene Research Products, San Diego, CA) as described ( ).

    Techniques: Activity Assay, Luciferase, Transfection, Soft Agar Assay, Cell Culture

    HER2 gene-amplified but not HER2-negative breast cancer cells are sensitive to FASN inhibition-induced apoptosis Top. HER2 protein expression in a panel of human breast cancer cell lines. Bottom. Fold-change in the C75-induced percentage of sub-G 1 , G 0 /G 1 , S, and G 2 /M phases relative to untreated MDA-MB-231 and BT-474 cells (=1-fold in each phase). Cell cycle analyses were repeated at least three times. Standard deviations were less 0.1-fold for each experimental condition.

    Journal: Histology and histopathology

    Article Title: Clinical and therapeutic relevance of the metabolic oncogene fatty acid synthase in HER2+ breast cancer

    doi: 10.14670/HH-11-830

    Figure Lengend Snippet: HER2 gene-amplified but not HER2-negative breast cancer cells are sensitive to FASN inhibition-induced apoptosis Top. HER2 protein expression in a panel of human breast cancer cell lines. Bottom. Fold-change in the C75-induced percentage of sub-G 1 , G 0 /G 1 , S, and G 2 /M phases relative to untreated MDA-MB-231 and BT-474 cells (=1-fold in each phase). Cell cycle analyses were repeated at least three times. Standard deviations were less 0.1-fold for each experimental condition.

    Article Snippet: HER2 expression was assesed by western blotting using a monoclonal antibody against HER2 (Ab-3 clone, Oncogene Research Products, San Diego, CA) as described ( ).

    Techniques: Amplification, Inhibition, Expressing, Multiple Displacement Amplification

    Immunophenotypic classification of FASN-overexpression in HER2-negative and HER2-positive invasive breast carcinomas.

    Journal: Histology and histopathology

    Article Title: Clinical and therapeutic relevance of the metabolic oncogene fatty acid synthase in HER2+ breast cancer

    doi: 10.14670/HH-11-830

    Figure Lengend Snippet: Immunophenotypic classification of FASN-overexpression in HER2-negative and HER2-positive invasive breast carcinomas.

    Article Snippet: HER2 expression was assesed by western blotting using a monoclonal antibody against HER2 (Ab-3 clone, Oncogene Research Products, San Diego, CA) as described ( ).

    Techniques: Over Expression

    Sulforaphane inhibits multiple cancer development-associated signaling pathways in A2780 cells. (A) Effect of sulforaphane on intracellular levels of Her2 through immunofluorescence assays (magnification, ×100). (B) Western blot assays were performed to examine the effect of sulforaphane on the 10 cancer-associated signaling pathway reporter activities, including Bcl-2, Bax, Cyto-c, Caspase-3, p-AKT, p-NF-κB, P53, P27, cyclin-D1 and cMyc. The values are presented as the mean ± standard deviation (n=8-10) of the samples. + P

    Journal: International Journal of Molecular Medicine

    Article Title: Sulforaphane regulates apoptosis- and proliferation-related signaling pathways and synergizes with cisplatin to suppress human ovarian cancer

    doi: 10.3892/ijmm.2018.3860

    Figure Lengend Snippet: Sulforaphane inhibits multiple cancer development-associated signaling pathways in A2780 cells. (A) Effect of sulforaphane on intracellular levels of Her2 through immunofluorescence assays (magnification, ×100). (B) Western blot assays were performed to examine the effect of sulforaphane on the 10 cancer-associated signaling pathway reporter activities, including Bcl-2, Bax, Cyto-c, Caspase-3, p-AKT, p-NF-κB, P53, P27, cyclin-D1 and cMyc. The values are presented as the mean ± standard deviation (n=8-10) of the samples. + P

    Article Snippet: The tumor tissue samples were incubated with primary monoclonal antibodies against Her2 and KI67 (1:200; 9449; Cell Signaling Technology, Inc.) at 4°C overnight.

    Techniques: Immunofluorescence, Western Blot, Standard Deviation

    Sulforaphane effectively suppresses tumor growth and progression in the xenograft model of human ovarian cancer cells. (A) Tumor xenograft model. The A2780 cells were injected into the hindlimbs of nude mice; and (B) tumor size and (C) weight were observed and measured. (D) Analysis of TUNEL, KI67 and Her2 in cancer tissues by immunohistochemistry. A brown signal was considered positive staining for TUNEL, KI67 and Her2 (magnification, ×100). Arrows indicate representative positive cells. (E) Reverse transcription-quantitative polymerase chain reaction assays were used to examine the effect of sulforaphane on the seven cancer-associated signaling pathway reporter activities, including Bcl-2, Bax, P53, P27, Cyclin-D1, cMyc and Her2. The values are presented as the mean ± standard deviation (n=8-10) of the samples. + P

    Journal: International Journal of Molecular Medicine

    Article Title: Sulforaphane regulates apoptosis- and proliferation-related signaling pathways and synergizes with cisplatin to suppress human ovarian cancer

    doi: 10.3892/ijmm.2018.3860

    Figure Lengend Snippet: Sulforaphane effectively suppresses tumor growth and progression in the xenograft model of human ovarian cancer cells. (A) Tumor xenograft model. The A2780 cells were injected into the hindlimbs of nude mice; and (B) tumor size and (C) weight were observed and measured. (D) Analysis of TUNEL, KI67 and Her2 in cancer tissues by immunohistochemistry. A brown signal was considered positive staining for TUNEL, KI67 and Her2 (magnification, ×100). Arrows indicate representative positive cells. (E) Reverse transcription-quantitative polymerase chain reaction assays were used to examine the effect of sulforaphane on the seven cancer-associated signaling pathway reporter activities, including Bcl-2, Bax, P53, P27, Cyclin-D1, cMyc and Her2. The values are presented as the mean ± standard deviation (n=8-10) of the samples. + P

    Article Snippet: The tumor tissue samples were incubated with primary monoclonal antibodies against Her2 and KI67 (1:200; 9449; Cell Signaling Technology, Inc.) at 4°C overnight.

    Techniques: Injection, Mouse Assay, TUNEL Assay, Immunohistochemistry, Staining, Real-time Polymerase Chain Reaction, Standard Deviation

    The HSF1 transcriptional program is impaired by HER2 inhibition. ( a ) Schematic overview of HSF1-regulated gene expression. Transcriptionally activated and -repressed target genes analyzed here are indicated. ( b and c ) HER2 inhibition impairs the HSF1-dependent inducible heat-shock response ( b ) and the tumor-promoting ( c ) transcriptional program of HSF1. SK-BR-3 cells were treated with 2 μ M CP724.714 or DMSO for 48 h and mRNA was isolated. qRT-PCR of Hsp70 (* P =0.0396), Hsp110 (*** P =0.001), Hsp90 α (** P =0.0049), SPTAN (* P =0.0362), CDC6 (*** P =0.001), FASN (** P =0.0069) and CBX3 ( P =0.067), each normalized to 36B4 mRNA. Relative values are given in (ratio (2 −ddCT )). Error bars indicate ±S.E.M. of three independent experiments, repeated twice each with all in triplicates. Student's t -test, two-tailed, P- value: * P

    Journal: Cell Death & Disease

    Article Title: HER2/ErbB2 activates HSF1 and thereby controls HSP90 clients including MIF in HER2-overexpressing breast cancer

    doi: 10.1038/cddis.2013.508

    Figure Lengend Snippet: The HSF1 transcriptional program is impaired by HER2 inhibition. ( a ) Schematic overview of HSF1-regulated gene expression. Transcriptionally activated and -repressed target genes analyzed here are indicated. ( b and c ) HER2 inhibition impairs the HSF1-dependent inducible heat-shock response ( b ) and the tumor-promoting ( c ) transcriptional program of HSF1. SK-BR-3 cells were treated with 2 μ M CP724.714 or DMSO for 48 h and mRNA was isolated. qRT-PCR of Hsp70 (* P =0.0396), Hsp110 (*** P =0.001), Hsp90 α (** P =0.0049), SPTAN (* P =0.0362), CDC6 (*** P =0.001), FASN (** P =0.0069) and CBX3 ( P =0.067), each normalized to 36B4 mRNA. Relative values are given in (ratio (2 −ddCT )). Error bars indicate ±S.E.M. of three independent experiments, repeated twice each with all in triplicates. Student's t -test, two-tailed, P- value: * P

    Article Snippet: Primary antibodies against HER2 (4B5, Ventana Medical Systems Inc.) and phospho-Ser326 HSF1 (EP1713Y, Abcam) and OptiView DAB IHC detection kits (Ventana Medical Systems Inc.) were used.

    Techniques: Inhibition, Expressing, Isolation, Quantitative RT-PCR, Two Tailed Test

    Blocking the HER2–PI3K–mTOR pathway reduces MIF protein levels. ( a and b ) Inhibition of the HER2–PI3K axis, but not the HER2–MEK–ERK1/2 axis, destabilizes MIF protein. SK-BR-3 cells were treated with 1 μ M CP724.714 (specific HER2 inhibitor), 25 μ M Ly294002 (PI3K inhibitor) or 10 μ M U0126 (MEK1/2 inhibitor) ( a ), or with 1 μ M CP724.714, 10 μ M U0126 or 50 μ M zVAD (panCaspase inhibitor) ( b ) alone or in combination as indicated. Immunoblot analyses. pERK1/2 and pAKT staining serve as positive control for respective inhibition. Gapdh, loading control. ( c ) Inhibition of mTOR destabilizes MIF protein. HER2-overexpressing BT-474, MDA-MB-453 and SK-BR-3 cells were treated with 50 nM of Rapamycin for 48 h or left untreated. Immunoblot analyses. pS6 serves as functional control of mTOR inhibition. Actin, loading control. ( d ) The dual HER2/EGFR inhibitor Lapatinib markedly reduces MIF levels. SK-BR-3 cells were treated or not with Lapatinib for 48 h. Immunoblot analysis. pERK1/2 is a functional control for HER2 inhibition. Gapdh, loading control. ( e ) Lapatinib blocks survival. SK-BR-3 cells were seeded (day 0) and cultured for 24 h (day 1). Cells were then treated or left untreated with 2 μ M Lapatinib for 48 h and followed up to day 5. Confluence measured daily by CELIGO Cytometer. Error bars indicate the S.E.M. of a triplicate experiment. Student's t -test of day 4, two-tailed, P- value: *** P

    Journal: Cell Death & Disease

    Article Title: HER2/ErbB2 activates HSF1 and thereby controls HSP90 clients including MIF in HER2-overexpressing breast cancer

    doi: 10.1038/cddis.2013.508

    Figure Lengend Snippet: Blocking the HER2–PI3K–mTOR pathway reduces MIF protein levels. ( a and b ) Inhibition of the HER2–PI3K axis, but not the HER2–MEK–ERK1/2 axis, destabilizes MIF protein. SK-BR-3 cells were treated with 1 μ M CP724.714 (specific HER2 inhibitor), 25 μ M Ly294002 (PI3K inhibitor) or 10 μ M U0126 (MEK1/2 inhibitor) ( a ), or with 1 μ M CP724.714, 10 μ M U0126 or 50 μ M zVAD (panCaspase inhibitor) ( b ) alone or in combination as indicated. Immunoblot analyses. pERK1/2 and pAKT staining serve as positive control for respective inhibition. Gapdh, loading control. ( c ) Inhibition of mTOR destabilizes MIF protein. HER2-overexpressing BT-474, MDA-MB-453 and SK-BR-3 cells were treated with 50 nM of Rapamycin for 48 h or left untreated. Immunoblot analyses. pS6 serves as functional control of mTOR inhibition. Actin, loading control. ( d ) The dual HER2/EGFR inhibitor Lapatinib markedly reduces MIF levels. SK-BR-3 cells were treated or not with Lapatinib for 48 h. Immunoblot analysis. pERK1/2 is a functional control for HER2 inhibition. Gapdh, loading control. ( e ) Lapatinib blocks survival. SK-BR-3 cells were seeded (day 0) and cultured for 24 h (day 1). Cells were then treated or left untreated with 2 μ M Lapatinib for 48 h and followed up to day 5. Confluence measured daily by CELIGO Cytometer. Error bars indicate the S.E.M. of a triplicate experiment. Student's t -test of day 4, two-tailed, P- value: *** P

    Article Snippet: Primary antibodies against HER2 (4B5, Ventana Medical Systems Inc.) and phospho-Ser326 HSF1 (EP1713Y, Abcam) and OptiView DAB IHC detection kits (Ventana Medical Systems Inc.) were used.

    Techniques: Blocking Assay, Inhibition, Staining, Positive Control, Multiple Displacement Amplification, Functional Assay, Cell Culture, Cytometry, Two Tailed Test

    HER2 inhibition leads to HSF1 inactivation and subsequent inactivation of the HSP90 chaperone. ( a and b ) Inhibition of HER2 inactivates HSF1 ( a ) and thereby destabilizes HSP90 clients ( b ). SK-BR-3 cells were treated with 2 μ M CP724.714 for 48 h. Protein levels of pSer 326 HSF1, total HSF1, Hsp90 α , Hsp70 and Hsp27 ( a ) and MIF, AKT, Bcl-xl and mutant p53 R175H ( b ) were assessed by immunoblots. Gapdh, loading control. ( c ) Inhibition of the HER2–PI3K axis, but not the HER2–ERK1/2 axis, inactivates HSF1 and downstream chaperones. SK-BR-3 cells were treated with 1 μ M CP724.714 (specific HER2 inhibitor), 25 μ M Ly294002 (PI3K inhibitor) or 10 μ M U0126 (MEK inhibitor) alone or in combination as indicated. Immunoblot analysis. Arrow indicates unrelated band. Hsc70, loading control. ( d ) Inhibition of mTOR prevents HSF1 activation. Cells were treated or left untreated with 50 nM Rapamycin for 48 h. Immunoblot analyses. pS6 serves as functional control of mTOR inhibition. Actin, loading control. ( e ) HSF1 silencing destabilizes MIF protein. MDA-MB-453 and MDA-MB-231 cells were transfected with different siRNAs against HSF1 for 3 days. Cell lysates were immunoblotted for MIF, panHsp90, Hsp70, Bcl-xl and c-Raf. Representative blots are shown. Actin, loading control. Arrow indicates related band. ( f ) The heat-shock response is impaired in response to inhibition of the HER2 pathway. SK-BR-3 cells were left untreated or treated with 2 μ M CP724.714 or 25 μ M Ly294002 for 48 h, followed by heat shock at 42 °C for 1 h where indicated. Protein lysates were prepared after a recovery of 3 h. Immunoblot analysis for pSer 326 HSF1 and total HSF1. Note that the HS-induced hyperphosphorylated species of total HSF1 (slower migrating band) is blocked by HER2 and PI3K inhibition. pS6 serves as control for HER2 and PI3K inhibition. Actin, loading control. ( g ) The heat-shock response is markedly attenuated in response to HER2 inhibition. SK-BR-3 were treated with 2 μ M CP724.714 or 25 μ M Ly294002 for 48 h. After heat shock at 42 °C for 1 h followed by a 3-h recovery, cells were immunostained for total HSF1 with DAPI counterstain. Top, representative immunofluorescence of heat-shocked control cells and heat-shocked/CP274.714-treated cells. Arrows indicate HSF1-positive nuclei with a granular nuclear pattern. Only those were counted. Bottom, quantification of HSF1-positive nuclei. For every sample, eight random fields ( × 20 magnification) in duplicates were counted. The number of HSF1-positive cells as percent of total nuclei (DAPI stained) is shown. Error bars indicates ±S.E.M. Student's t -test, two-tailed, P- value: *** P

    Journal: Cell Death & Disease

    Article Title: HER2/ErbB2 activates HSF1 and thereby controls HSP90 clients including MIF in HER2-overexpressing breast cancer

    doi: 10.1038/cddis.2013.508

    Figure Lengend Snippet: HER2 inhibition leads to HSF1 inactivation and subsequent inactivation of the HSP90 chaperone. ( a and b ) Inhibition of HER2 inactivates HSF1 ( a ) and thereby destabilizes HSP90 clients ( b ). SK-BR-3 cells were treated with 2 μ M CP724.714 for 48 h. Protein levels of pSer 326 HSF1, total HSF1, Hsp90 α , Hsp70 and Hsp27 ( a ) and MIF, AKT, Bcl-xl and mutant p53 R175H ( b ) were assessed by immunoblots. Gapdh, loading control. ( c ) Inhibition of the HER2–PI3K axis, but not the HER2–ERK1/2 axis, inactivates HSF1 and downstream chaperones. SK-BR-3 cells were treated with 1 μ M CP724.714 (specific HER2 inhibitor), 25 μ M Ly294002 (PI3K inhibitor) or 10 μ M U0126 (MEK inhibitor) alone or in combination as indicated. Immunoblot analysis. Arrow indicates unrelated band. Hsc70, loading control. ( d ) Inhibition of mTOR prevents HSF1 activation. Cells were treated or left untreated with 50 nM Rapamycin for 48 h. Immunoblot analyses. pS6 serves as functional control of mTOR inhibition. Actin, loading control. ( e ) HSF1 silencing destabilizes MIF protein. MDA-MB-453 and MDA-MB-231 cells were transfected with different siRNAs against HSF1 for 3 days. Cell lysates were immunoblotted for MIF, panHsp90, Hsp70, Bcl-xl and c-Raf. Representative blots are shown. Actin, loading control. Arrow indicates related band. ( f ) The heat-shock response is impaired in response to inhibition of the HER2 pathway. SK-BR-3 cells were left untreated or treated with 2 μ M CP724.714 or 25 μ M Ly294002 for 48 h, followed by heat shock at 42 °C for 1 h where indicated. Protein lysates were prepared after a recovery of 3 h. Immunoblot analysis for pSer 326 HSF1 and total HSF1. Note that the HS-induced hyperphosphorylated species of total HSF1 (slower migrating band) is blocked by HER2 and PI3K inhibition. pS6 serves as control for HER2 and PI3K inhibition. Actin, loading control. ( g ) The heat-shock response is markedly attenuated in response to HER2 inhibition. SK-BR-3 were treated with 2 μ M CP724.714 or 25 μ M Ly294002 for 48 h. After heat shock at 42 °C for 1 h followed by a 3-h recovery, cells were immunostained for total HSF1 with DAPI counterstain. Top, representative immunofluorescence of heat-shocked control cells and heat-shocked/CP274.714-treated cells. Arrows indicate HSF1-positive nuclei with a granular nuclear pattern. Only those were counted. Bottom, quantification of HSF1-positive nuclei. For every sample, eight random fields ( × 20 magnification) in duplicates were counted. The number of HSF1-positive cells as percent of total nuclei (DAPI stained) is shown. Error bars indicates ±S.E.M. Student's t -test, two-tailed, P- value: *** P

    Article Snippet: Primary antibodies against HER2 (4B5, Ventana Medical Systems Inc.) and phospho-Ser326 HSF1 (EP1713Y, Abcam) and OptiView DAB IHC detection kits (Ventana Medical Systems Inc.) were used.

    Techniques: Inhibition, Mutagenesis, Western Blot, Activation Assay, Functional Assay, Multiple Displacement Amplification, Transfection, Immunofluorescence, Staining, Two Tailed Test

    Specific inhibition of HER2 signaling in HER2-overexpressing breast cancer cells reduces MIF protein levels. ( a ) Endogenous HER2 protein levels in human breast cancer cells. Representative immunoblot of cell lysates from the indicated cell lines. mErbB2, murine ErbB2 cell line. Actin, loading control. ( b ) HER2 inhibition destabilizes endogenous MIF protein. The indicated cells were treated with 2 μ M CP724.714 or DMSO for 48 h. Immunoblot. Actin, loading control. ( c and d ) The specific HER2 inhibitor CP724.714 reduces endogenous MIF levels in a dose- ( c ) and time- ( d ) dependent manner. MDA-MB-231 serves as negative control. pAKT and pERK1/2 are functional controls for HER2 inhibition. Immunoblot analyses, Gapdh as loading control. ( e ) Depletion of HER2 in HER2-overexpressing cells leads to reduced MIF levels. SK-BR-3 cells were transfected with two different siRNA against HER2 (1 and 3) or control siRNA (scr). After 3 days, protein levels were assessed by immunoblots. Gapdh, loading control. ( f ) In contrast to reduced MIF protein, corresponding MIF mRNA levels remain unchanged after CP724.714 treatment (2 μM). SK-BR-3 cells, qRT-PCR normalized to 36B4. Relative values in (ratio (2 −ddCT )). Error bars indicate S.E.M. of two independent experiments in triplicates each. ( g ) HER2 inhibition causes growth inhibition in HER2-overexpressing cells. Cells were seeded (day 0) and cultured for 24 h (day 1), then treated with 2 μ M of CP724.714 (CP) for 24 h or left untreated, and followed up to day 5. Cell confluence measured daily by CELIGO Cytometer for MDA-MB-453 (*** P =0.0006) and SK-BR-3 (** P =0.0037). Error bars indicate the ±S.E.M of two independent experiments in duplicates each. Student's t -test of day 5, one-tailed, P- value: * P

    Journal: Cell Death & Disease

    Article Title: HER2/ErbB2 activates HSF1 and thereby controls HSP90 clients including MIF in HER2-overexpressing breast cancer

    doi: 10.1038/cddis.2013.508

    Figure Lengend Snippet: Specific inhibition of HER2 signaling in HER2-overexpressing breast cancer cells reduces MIF protein levels. ( a ) Endogenous HER2 protein levels in human breast cancer cells. Representative immunoblot of cell lysates from the indicated cell lines. mErbB2, murine ErbB2 cell line. Actin, loading control. ( b ) HER2 inhibition destabilizes endogenous MIF protein. The indicated cells were treated with 2 μ M CP724.714 or DMSO for 48 h. Immunoblot. Actin, loading control. ( c and d ) The specific HER2 inhibitor CP724.714 reduces endogenous MIF levels in a dose- ( c ) and time- ( d ) dependent manner. MDA-MB-231 serves as negative control. pAKT and pERK1/2 are functional controls for HER2 inhibition. Immunoblot analyses, Gapdh as loading control. ( e ) Depletion of HER2 in HER2-overexpressing cells leads to reduced MIF levels. SK-BR-3 cells were transfected with two different siRNA against HER2 (1 and 3) or control siRNA (scr). After 3 days, protein levels were assessed by immunoblots. Gapdh, loading control. ( f ) In contrast to reduced MIF protein, corresponding MIF mRNA levels remain unchanged after CP724.714 treatment (2 μM). SK-BR-3 cells, qRT-PCR normalized to 36B4. Relative values in (ratio (2 −ddCT )). Error bars indicate S.E.M. of two independent experiments in triplicates each. ( g ) HER2 inhibition causes growth inhibition in HER2-overexpressing cells. Cells were seeded (day 0) and cultured for 24 h (day 1), then treated with 2 μ M of CP724.714 (CP) for 24 h or left untreated, and followed up to day 5. Cell confluence measured daily by CELIGO Cytometer for MDA-MB-453 (*** P =0.0006) and SK-BR-3 (** P =0.0037). Error bars indicate the ±S.E.M of two independent experiments in duplicates each. Student's t -test of day 5, one-tailed, P- value: * P

    Article Snippet: Primary antibodies against HER2 (4B5, Ventana Medical Systems Inc.) and phospho-Ser326 HSF1 (EP1713Y, Abcam) and OptiView DAB IHC detection kits (Ventana Medical Systems Inc.) were used.

    Techniques: Inhibition, Multiple Displacement Amplification, Negative Control, Functional Assay, Transfection, Western Blot, Quantitative RT-PCR, Cell Culture, Cytometry, One-tailed Test

    In human HER2-positive breast cancers, HER2 levels correlate with pSer326 HSF1 activity. ( a and b ) Correlation in staining intensity between HER2 and activated HSF1 (pSer326) in human breast cancer. ( a ) Quantitative immunohistochemistry of HER2 and pSer326 HSF1 protein in HER2-amplified invasive ductal carcinoma with 2+ and 3+ HER2 staining intensity, respectively. Two regions encompassing > 60 cells were quantified per case; DAB color intensity displayed as values on an inverted 8 bit gray scale, 0=white, 255=black. ( b ) Representative photomicrographs of cases of invasive ductal carcinoma quantified in a . Immunohistochemical staining for HER2 and pSer326 HSF1. DAB with hematoxylin counterstain, × 400 magnification. Bar, 100 μ m. ( c ) Proposed model summarizing the findings of this study. In HER2-overexpressing cells, the PI3K–AKT–mTOR pathway is the main signaling axis that leads to phosphorylation of HSF1 at Ser326, which activates HSF1 transcriptional activity and induces, among other target genes, expression of heat-shock proteins. The activated HSP90 machinery stabilizes a broad panel of oncogenic and tumor-promoting proteins. Dashed lines mean feedback loop

    Journal: Cell Death & Disease

    Article Title: HER2/ErbB2 activates HSF1 and thereby controls HSP90 clients including MIF in HER2-overexpressing breast cancer

    doi: 10.1038/cddis.2013.508

    Figure Lengend Snippet: In human HER2-positive breast cancers, HER2 levels correlate with pSer326 HSF1 activity. ( a and b ) Correlation in staining intensity between HER2 and activated HSF1 (pSer326) in human breast cancer. ( a ) Quantitative immunohistochemistry of HER2 and pSer326 HSF1 protein in HER2-amplified invasive ductal carcinoma with 2+ and 3+ HER2 staining intensity, respectively. Two regions encompassing > 60 cells were quantified per case; DAB color intensity displayed as values on an inverted 8 bit gray scale, 0=white, 255=black. ( b ) Representative photomicrographs of cases of invasive ductal carcinoma quantified in a . Immunohistochemical staining for HER2 and pSer326 HSF1. DAB with hematoxylin counterstain, × 400 magnification. Bar, 100 μ m. ( c ) Proposed model summarizing the findings of this study. In HER2-overexpressing cells, the PI3K–AKT–mTOR pathway is the main signaling axis that leads to phosphorylation of HSF1 at Ser326, which activates HSF1 transcriptional activity and induces, among other target genes, expression of heat-shock proteins. The activated HSP90 machinery stabilizes a broad panel of oncogenic and tumor-promoting proteins. Dashed lines mean feedback loop

    Article Snippet: Primary antibodies against HER2 (4B5, Ventana Medical Systems Inc.) and phospho-Ser326 HSF1 (EP1713Y, Abcam) and OptiView DAB IHC detection kits (Ventana Medical Systems Inc.) were used.

    Techniques: Activity Assay, Staining, Immunohistochemistry, Amplification, Expressing

    Comparison of gene amplification patterns of Her-2, EGFR and cyclin D1 in primary tumor and the corresponding metastatic lymph node (LN) (×400). High-level amplification/high-level amplification of Her-2 in primary tumor ( A ) and LN ( B ); high-level amplification/no amplification of Her-2 in primary tumor ( C ) and LN ( D ); high-level amplification/no amplification of EGFR in primary tumor ( E ) and LN ( F ); low-level amplification/low-level amplification of EGFR in primary tumor ( G ) and LN ( H ); high-level amplification/no amplification of cyclin D1 in primary tumor ( I ) and LN ( J ); high-level amplification/high-level amplification of cyclin D1 in primary tumor ( K ) and LN ( L ).

    Journal: Journal of Korean Medical Science

    Article Title: Comparison of Her-2, EGFR and Cyclin D1 in Primary Breast Cancer and Paired Metastatic Lymph Nodes: An Immunohistochemical and Chromogenic In Situ Hybridization Study

    doi: 10.3346/jkms.2008.23.6.1053

    Figure Lengend Snippet: Comparison of gene amplification patterns of Her-2, EGFR and cyclin D1 in primary tumor and the corresponding metastatic lymph node (LN) (×400). High-level amplification/high-level amplification of Her-2 in primary tumor ( A ) and LN ( B ); high-level amplification/no amplification of Her-2 in primary tumor ( C ) and LN ( D ); high-level amplification/no amplification of EGFR in primary tumor ( E ) and LN ( F ); low-level amplification/low-level amplification of EGFR in primary tumor ( G ) and LN ( H ); high-level amplification/no amplification of cyclin D1 in primary tumor ( I ) and LN ( J ); high-level amplification/high-level amplification of cyclin D1 in primary tumor ( K ) and LN ( L ).

    Article Snippet: Immunohistochemistry After deparaffinization and rehydration, 4-µm thick sections on silane-coated slides were heat-pretreated in a citrate buffer (pH 7.3 at 92℃ in microwave oven) and examined by immunostaining using specific antibodies against Her-2 (CB11; Novocastra Laboratories, Newcastle, U.K.), EGFR (Novocastra Laboratories), and cyclin D1 (P2D11F11; Novocastra Laboratories).

    Techniques: Amplification

    Comparison of immunostaining patterns of Her-2, EGFR and cyclin D1 in primary tumor and the corresponding metastatic lymph node (LN) (×200). 3+/3+ expression of Her-2 in primary tumor ( A ) and LN ( B ); 2+/- expression of Her-2 in primary tumor ( C ) and LN ( D ); 1+/2+ expression of Her-2 in primary tumor ( E ) and LN ( F ), 3+/- expression of EGFR in primary tumor ( G ) and LN ( H ); -/2+ expression of EGFR in primary tumor ( I ) and LN ( J ); -/3+ expression of cyclin D1 in primary tumor ( K ) and LN ( L ).

    Journal: Journal of Korean Medical Science

    Article Title: Comparison of Her-2, EGFR and Cyclin D1 in Primary Breast Cancer and Paired Metastatic Lymph Nodes: An Immunohistochemical and Chromogenic In Situ Hybridization Study

    doi: 10.3346/jkms.2008.23.6.1053

    Figure Lengend Snippet: Comparison of immunostaining patterns of Her-2, EGFR and cyclin D1 in primary tumor and the corresponding metastatic lymph node (LN) (×200). 3+/3+ expression of Her-2 in primary tumor ( A ) and LN ( B ); 2+/- expression of Her-2 in primary tumor ( C ) and LN ( D ); 1+/2+ expression of Her-2 in primary tumor ( E ) and LN ( F ), 3+/- expression of EGFR in primary tumor ( G ) and LN ( H ); -/2+ expression of EGFR in primary tumor ( I ) and LN ( J ); -/3+ expression of cyclin D1 in primary tumor ( K ) and LN ( L ).

    Article Snippet: Immunohistochemistry After deparaffinization and rehydration, 4-µm thick sections on silane-coated slides were heat-pretreated in a citrate buffer (pH 7.3 at 92℃ in microwave oven) and examined by immunostaining using specific antibodies against Her-2 (CB11; Novocastra Laboratories, Newcastle, U.K.), EGFR (Novocastra Laboratories), and cyclin D1 (P2D11F11; Novocastra Laboratories).

    Techniques: Immunostaining, Expressing

    Representative pictures of HER-2 IHC and FISH staining on models EC004, 016, 039, 044 and 054 with matched human primary tumor tissues. HER-2 moderate staining (++) and FISH GCN increase were detected in models EC039 and EC044 and matched primary tumor tissues. EC004, EC016 and EC054 were shown to be HER-2 IHC negative and FISH non-AMP.

    Journal: Journal of Translational Medicine

    Article Title: Trastuzumab anti-tumor efficacy in patient-derived esophageal squamous cell carcinoma xenograft (PDECX) mouse models

    doi: 10.1186/1479-5876-10-180

    Figure Lengend Snippet: Representative pictures of HER-2 IHC and FISH staining on models EC004, 016, 039, 044 and 054 with matched human primary tumor tissues. HER-2 moderate staining (++) and FISH GCN increase were detected in models EC039 and EC044 and matched primary tumor tissues. EC004, EC016 and EC054 were shown to be HER-2 IHC negative and FISH non-AMP.

    Article Snippet: Antibodies against HER-2 (#06-562) was purchased from Upstate Biotechnology; p-Mtor-Ser 2448 (No. 2971) from DAKO Biotechnology, and mTor (No 2972), AKT (No 9272), p44/42 MAP Kinase (No 9102), S6 (No 2317), 4EBP1 (No 9452) and p-AKT-Ser 473 (M3628), Phospho-p44/42-MAPK-Thr202/Tyr 204 (No 4370), p-S6-Ser 240/244 (No 2215), p-4EBP1-Thr 70 (No 9455), and anti-GAPDH were purchased from CST Biotechnology.

    Techniques: Immunohistochemistry, Fluorescence In Situ Hybridization, Staining

    Microscopic analysis of the HER2-ECD-induced HER2-negative cells treated with Tra-IR700-mediated PIT. A, morphological changes of the tumor cells after PIT. MKN1 and MKN45 were cultured with Ad/HER2-ECD for 48 h and were treated with Tra-IR700 for 6 h.

    Journal: Molecular cancer therapeutics

    Article Title: Trastuzumab-based photoimmunotherapy integrated with viral HER2 transduction inhibits peritoneally disseminated HER2-negative cancer

    doi: 10.1158/1535-7163.MCT-15-0644

    Figure Lengend Snippet: Microscopic analysis of the HER2-ECD-induced HER2-negative cells treated with Tra-IR700-mediated PIT. A, morphological changes of the tumor cells after PIT. MKN1 and MKN45 were cultured with Ad/HER2-ECD for 48 h and were treated with Tra-IR700 for 6 h.

    Article Snippet: Membranes were incubated with primary antibodies against HER2-ECD (Thermo Scientific., Yokohama, Japan) overnight at 4°C and visualized using an Amersham ECL chemiluminescence system (GE Healthcare UK Ltd.) according to the manufacturer’s protocol.

    Techniques: Cell Culture

    HER2-ECD expression induced by Ad/HER2-ECD and the distribution of Tra-IR700 in gastric cancer cells. A, Western blot analysis revealed that MKN1 and MKN45 did not express wild-type (wt) HER2 (185 kDa), whereas N87 cells expressed it. Infection with Ad/HER2-ECD

    Journal: Molecular cancer therapeutics

    Article Title: Trastuzumab-based photoimmunotherapy integrated with viral HER2 transduction inhibits peritoneally disseminated HER2-negative cancer

    doi: 10.1158/1535-7163.MCT-15-0644

    Figure Lengend Snippet: HER2-ECD expression induced by Ad/HER2-ECD and the distribution of Tra-IR700 in gastric cancer cells. A, Western blot analysis revealed that MKN1 and MKN45 did not express wild-type (wt) HER2 (185 kDa), whereas N87 cells expressed it. Infection with Ad/HER2-ECD

    Article Snippet: Membranes were incubated with primary antibodies against HER2-ECD (Thermo Scientific., Yokohama, Japan) overnight at 4°C and visualized using an Amersham ECL chemiluminescence system (GE Healthcare UK Ltd.) according to the manufacturer’s protocol.

    Techniques: Expressing, Western Blot, Infection

    Evaluation of the HER2 expression induced by Ad/HER2-ECD on the tumor cells and normal mice peritoneal cells ex vivo . A, a culture of normal mouse peritoneal cells (2×10 5 cells) and the co-culture of normal mouse peritoneal cells and MKN45 cells

    Journal: Molecular cancer therapeutics

    Article Title: Trastuzumab-based photoimmunotherapy integrated with viral HER2 transduction inhibits peritoneally disseminated HER2-negative cancer

    doi: 10.1158/1535-7163.MCT-15-0644

    Figure Lengend Snippet: Evaluation of the HER2 expression induced by Ad/HER2-ECD on the tumor cells and normal mice peritoneal cells ex vivo . A, a culture of normal mouse peritoneal cells (2×10 5 cells) and the co-culture of normal mouse peritoneal cells and MKN45 cells

    Article Snippet: Membranes were incubated with primary antibodies against HER2-ECD (Thermo Scientific., Yokohama, Japan) overnight at 4°C and visualized using an Amersham ECL chemiluminescence system (GE Healthcare UK Ltd.) according to the manufacturer’s protocol.

    Techniques: Expressing, Mouse Assay, Ex Vivo, Co-Culture Assay

    Inhibitory effects by the integrated therapy of Ad/HER2-ECD and Tra-IR700–mediated PIT in vivo . A, mice peritoneally bearing MKN45 cells were treated with or without Ad/HER2-ECD (1×10 8 pfu) and/or Tra-IR700 (80 µg) on days 5 and

    Journal: Molecular cancer therapeutics

    Article Title: Trastuzumab-based photoimmunotherapy integrated with viral HER2 transduction inhibits peritoneally disseminated HER2-negative cancer

    doi: 10.1158/1535-7163.MCT-15-0644

    Figure Lengend Snippet: Inhibitory effects by the integrated therapy of Ad/HER2-ECD and Tra-IR700–mediated PIT in vivo . A, mice peritoneally bearing MKN45 cells were treated with or without Ad/HER2-ECD (1×10 8 pfu) and/or Tra-IR700 (80 µg) on days 5 and

    Article Snippet: Membranes were incubated with primary antibodies against HER2-ECD (Thermo Scientific., Yokohama, Japan) overnight at 4°C and visualized using an Amersham ECL chemiluminescence system (GE Healthcare UK Ltd.) according to the manufacturer’s protocol.

    Techniques: In Vivo, Mouse Assay