p erbb2 tyr1248  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p erbb2 tyr1248
    Primer sequences used for the qPCR analysis
    P Erbb2 Tyr1248, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Dysregulation of Neuregulin-1/ErbB signaling in the hippocampus of rats after administration of doxorubicin"

    Article Title: Dysregulation of Neuregulin-1/ErbB signaling in the hippocampus of rats after administration of doxorubicin

    Journal: Drug Design, Development and Therapy

    doi: 10.2147/DDDT.S151511

    Primer sequences used for the qPCR analysis
    Figure Legend Snippet: Primer sequences used for the qPCR analysis

    Techniques Used: Amplification

    Effect of Dox on gene expression of ErbB2, ErbB4, and the ratio of pErbB4/ErbB4 and pErbB2/ErbB2 in the hippocampus. Notes: ErbB4 mRNA expression ( A ); pErbB4/ErbB4 ratio ( B ); ErbB2 mRNA expression ( C ); and pErbB2/ErbB2 ratio ( D ). Data are expressed as mean ± SEM (n=6–7). * p <0.05 and ** p <0.01 compared to the control group. Abbreviations: Dox, doxorubicin; DoxS, doxorubicin administration for short time; DoxL, doxorubicin administration for long time; SEM, standard error of the mean.
    Figure Legend Snippet: Effect of Dox on gene expression of ErbB2, ErbB4, and the ratio of pErbB4/ErbB4 and pErbB2/ErbB2 in the hippocampus. Notes: ErbB4 mRNA expression ( A ); pErbB4/ErbB4 ratio ( B ); ErbB2 mRNA expression ( C ); and pErbB2/ErbB2 ratio ( D ). Data are expressed as mean ± SEM (n=6–7). * p <0.05 and ** p <0.01 compared to the control group. Abbreviations: Dox, doxorubicin; DoxS, doxorubicin administration for short time; DoxL, doxorubicin administration for long time; SEM, standard error of the mean.

    Techniques Used: Expressing

    2247s  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc 2247s
    KEY RESOURCES TABLE
    2247s, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "KRAS G12C -independent feedback activation of wild-type RAS constrains KRAS G12C inhibitor efficacy"

    Article Title: KRAS G12C -independent feedback activation of wild-type RAS constrains KRAS G12C inhibitor efficacy

    Journal: Cell reports

    doi: 10.1016/j.celrep.2022.110993

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Derivative Assay, Recombinant, In Vitro, In Vivo, Software

    p erbb2 tyr1248  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p erbb2 tyr1248
    Primer sequences used for the qPCR analysis
    P Erbb2 Tyr1248, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Dysregulation of Neuregulin-1/ErbB signaling in the hippocampus of rats after administration of doxorubicin"

    Article Title: Dysregulation of Neuregulin-1/ErbB signaling in the hippocampus of rats after administration of doxorubicin

    Journal: Drug Design, Development and Therapy

    doi: 10.2147/DDDT.S151511

    Primer sequences used for the qPCR analysis
    Figure Legend Snippet: Primer sequences used for the qPCR analysis

    Techniques Used: Amplification

    Effect of Dox on gene expression of ErbB2, ErbB4, and the ratio of pErbB4/ErbB4 and pErbB2/ErbB2 in the hippocampus. Notes: ErbB4 mRNA expression ( A ); pErbB4/ErbB4 ratio ( B ); ErbB2 mRNA expression ( C ); and pErbB2/ErbB2 ratio ( D ). Data are expressed as mean ± SEM (n=6–7). * p <0.05 and ** p <0.01 compared to the control group. Abbreviations: Dox, doxorubicin; DoxS, doxorubicin administration for short time; DoxL, doxorubicin administration for long time; SEM, standard error of the mean.
    Figure Legend Snippet: Effect of Dox on gene expression of ErbB2, ErbB4, and the ratio of pErbB4/ErbB4 and pErbB2/ErbB2 in the hippocampus. Notes: ErbB4 mRNA expression ( A ); pErbB4/ErbB4 ratio ( B ); ErbB2 mRNA expression ( C ); and pErbB2/ErbB2 ratio ( D ). Data are expressed as mean ± SEM (n=6–7). * p <0.05 and ** p <0.01 compared to the control group. Abbreviations: Dox, doxorubicin; DoxS, doxorubicin administration for short time; DoxL, doxorubicin administration for long time; SEM, standard error of the mean.

    Techniques Used: Expressing

    anti phospho her2 tyr1248 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho her2 tyr1248 antibody
    Anti Phospho Her2 Tyr1248 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho her 2 tyr1248  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho her 2 tyr1248
    A–B, MdOS blocks <t>HER-2</t> phosphorylation and its downstream signal transduction in SK-OV-3 cells (A) and NIH-3T3/neu cells (B). C–D, Inhibition of EGF-stimulated EGFR phosphorylation and signal transduction by MdOS in A431 (C) and MDA-MB-468 (D) cells. Serum-starved cells were incubated with indicated concentrations of MdOS for 6 h at 37°C, EGF (50 ng/ml) was added to the cultures during the last 15-min treatment. E–F, Inhibition of VEGF-stimulated VEGFR2 phosphorylation and signal transduction by MdOS. HMEC (E) and NIH-3T3/flk-1 (F) cells were starved, then incubated with indicated concentrations of MdOS for 6 h at 37°C, VEGF (50 ng/ml) was added to the cultures during the last 15-min treatment. Protein samples were separated by SDS-PAGE and probed using the indicated antibodies. Representative data are shown.
    Phospho Her 2 Tyr1248, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "The Marine-Derived Oligosaccharide Sulfate (MdOS), a Novel Multiple Tyrosine Kinase Inhibitor, Combats Tumor Angiogenesis both In Vitro and In Vivo"

    Article Title: The Marine-Derived Oligosaccharide Sulfate (MdOS), a Novel Multiple Tyrosine Kinase Inhibitor, Combats Tumor Angiogenesis both In Vitro and In Vivo

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0003774

    A–B, MdOS blocks HER-2 phosphorylation and its downstream signal transduction in SK-OV-3 cells (A) and NIH-3T3/neu cells (B). C–D, Inhibition of EGF-stimulated EGFR phosphorylation and signal transduction by MdOS in A431 (C) and MDA-MB-468 (D) cells. Serum-starved cells were incubated with indicated concentrations of MdOS for 6 h at 37°C, EGF (50 ng/ml) was added to the cultures during the last 15-min treatment. E–F, Inhibition of VEGF-stimulated VEGFR2 phosphorylation and signal transduction by MdOS. HMEC (E) and NIH-3T3/flk-1 (F) cells were starved, then incubated with indicated concentrations of MdOS for 6 h at 37°C, VEGF (50 ng/ml) was added to the cultures during the last 15-min treatment. Protein samples were separated by SDS-PAGE and probed using the indicated antibodies. Representative data are shown.
    Figure Legend Snippet: A–B, MdOS blocks HER-2 phosphorylation and its downstream signal transduction in SK-OV-3 cells (A) and NIH-3T3/neu cells (B). C–D, Inhibition of EGF-stimulated EGFR phosphorylation and signal transduction by MdOS in A431 (C) and MDA-MB-468 (D) cells. Serum-starved cells were incubated with indicated concentrations of MdOS for 6 h at 37°C, EGF (50 ng/ml) was added to the cultures during the last 15-min treatment. E–F, Inhibition of VEGF-stimulated VEGFR2 phosphorylation and signal transduction by MdOS. HMEC (E) and NIH-3T3/flk-1 (F) cells were starved, then incubated with indicated concentrations of MdOS for 6 h at 37°C, VEGF (50 ng/ml) was added to the cultures during the last 15-min treatment. Protein samples were separated by SDS-PAGE and probed using the indicated antibodies. Representative data are shown.

    Techniques Used: Transduction, Inhibition, Incubation, SDS Page

    VEGFR2 was injected at the concentrations of 2.56, 0.64, 0.32, 0.16 and 0.08 µM (from top to bottom); HER-2 was injected at the concentrations of 0.53, 0.26, 0.13, 0.07 and 0.03 µM (from top to bottom); EGFR was injected at the concentrations of 6, 3, 1.5, 0.75, 0.38, 0.19 and 0.09 µM (from top to bottom); 6×his-tag was injected at the concentrations of 100, 10, 1, 0.1, 0.01 µM. Sensorgram responses at equilibrium were plotted against each concentration of compounds and the equilibrium dissociation constant (K D ) of the binding system was calculated using BIAeval software 3.1.
    Figure Legend Snippet: VEGFR2 was injected at the concentrations of 2.56, 0.64, 0.32, 0.16 and 0.08 µM (from top to bottom); HER-2 was injected at the concentrations of 0.53, 0.26, 0.13, 0.07 and 0.03 µM (from top to bottom); EGFR was injected at the concentrations of 6, 3, 1.5, 0.75, 0.38, 0.19 and 0.09 µM (from top to bottom); 6×his-tag was injected at the concentrations of 100, 10, 1, 0.1, 0.01 µM. Sensorgram responses at equilibrium were plotted against each concentration of compounds and the equilibrium dissociation constant (K D ) of the binding system was calculated using BIAeval software 3.1.

    Techniques Used: Injection, Concentration Assay, Binding Assay, Software

    VEGFR2 (A) and HER-2 (B) and EGFR (C) kinase assays were performed as described in in the presence of varying concentrations of ATP. Initial reaction velocity was expressed as the phosphorylation of poly(Glu, Tyr) 4∶1 substrate. All x, y data sets were multiplied by 100 for purposes of graphical presentation.
    Figure Legend Snippet: VEGFR2 (A) and HER-2 (B) and EGFR (C) kinase assays were performed as described in in the presence of varying concentrations of ATP. Initial reaction velocity was expressed as the phosphorylation of poly(Glu, Tyr) 4∶1 substrate. All x, y data sets were multiplied by 100 for purposes of graphical presentation.

    Techniques Used:

    pher2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pher2
    Pher2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    p her2 tyr1248  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p her2 tyr1248
    P Her2 Tyr1248, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    p neu  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p neu
    P Neu, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    p her2 erbb2 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p her2 erbb2 antibody
    a ) Representative Western blots showing levels of phosphorylated <t>erbB2</t> at Y877, Y1248, Y1248-a (which represents detection of Y1248 using an alternative antibody (p- erbB2-Antibody <t>(Tyr1248)/EGFR</t> (Tyr1173)) and Y12221/2 as well as total erbB2 (t-erbB2) and actin as a control protein in non-diabetic control hearts (C), diabetic hearts (D) and diabetic hearts chronically treated with AG825 (+AG825). b ) quantification of erbB2 expression relative to actin and c–f ) quantification of erbB2 phosphorylation at the stated tyrosine site relative to total erbB2 expression for all the groups studied by densitometry. N=4; * significantly different from control (p<0.05); ** significantly different from diabetes (p<0.05).
    P Her2 Erbb2 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Activation of EGFR/ERBB2 via Pathways Involving ERK1/2, P38 MAPK, AKT and FOXO Enhances Recovery of Diabetic Hearts from Ischemia-Reperfusion Injury"

    Article Title: Activation of EGFR/ERBB2 via Pathways Involving ERK1/2, P38 MAPK, AKT and FOXO Enhances Recovery of Diabetic Hearts from Ischemia-Reperfusion Injury

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0039066

    a ) Representative Western blots showing levels of phosphorylated erbB2 at Y877, Y1248, Y1248-a (which represents detection of Y1248 using an alternative antibody (p- erbB2-Antibody (Tyr1248)/EGFR (Tyr1173)) and Y12221/2 as well as total erbB2 (t-erbB2) and actin as a control protein in non-diabetic control hearts (C), diabetic hearts (D) and diabetic hearts chronically treated with AG825 (+AG825). b ) quantification of erbB2 expression relative to actin and c–f ) quantification of erbB2 phosphorylation at the stated tyrosine site relative to total erbB2 expression for all the groups studied by densitometry. N=4; * significantly different from control (p<0.05); ** significantly different from diabetes (p<0.05).
    Figure Legend Snippet: a ) Representative Western blots showing levels of phosphorylated erbB2 at Y877, Y1248, Y1248-a (which represents detection of Y1248 using an alternative antibody (p- erbB2-Antibody (Tyr1248)/EGFR (Tyr1173)) and Y12221/2 as well as total erbB2 (t-erbB2) and actin as a control protein in non-diabetic control hearts (C), diabetic hearts (D) and diabetic hearts chronically treated with AG825 (+AG825). b ) quantification of erbB2 expression relative to actin and c–f ) quantification of erbB2 phosphorylation at the stated tyrosine site relative to total erbB2 expression for all the groups studied by densitometry. N=4; * significantly different from control (p<0.05); ** significantly different from diabetes (p<0.05).

    Techniques Used: Western Blot, Expressing

    Panel a) are represenatative Western Blots following immunoprecipitations (IP) with either total-EGFR or total-erbB2 antibody and subsequent immunoblotting (IB) with both antibodies individually. Panel b) represents the mean ratio of erbB2/EGFR dimers as assessed by densitometry for non-diabetic control hearts (C), diabetic hearts (D) and diabetic hearts chronically treated with AG1478 (+AG1478) or AG825 (+AG825). N=4; * significantly different from control (p<0.05); ** significantly different from diabetes (p<0.05).
    Figure Legend Snippet: Panel a) are represenatative Western Blots following immunoprecipitations (IP) with either total-EGFR or total-erbB2 antibody and subsequent immunoblotting (IB) with both antibodies individually. Panel b) represents the mean ratio of erbB2/EGFR dimers as assessed by densitometry for non-diabetic control hearts (C), diabetic hearts (D) and diabetic hearts chronically treated with AG1478 (+AG1478) or AG825 (+AG825). N=4; * significantly different from control (p<0.05); ** significantly different from diabetes (p<0.05).

    Techniques Used: Western Blot

    Representative Western blots to show ischemia-induced changes in phosphorylation of EGFR and erbB2 receptors and downstream signaling molecules for a) normal (non-diabetic) control hearts (C) and those subjected to 40 min ischemia (CI) and b) diabetic hearts (D) and those subjected to 40 min ischemia (DI).
    Figure Legend Snippet: Representative Western blots to show ischemia-induced changes in phosphorylation of EGFR and erbB2 receptors and downstream signaling molecules for a) normal (non-diabetic) control hearts (C) and those subjected to 40 min ischemia (CI) and b) diabetic hearts (D) and those subjected to 40 min ischemia (DI).

    Techniques Used: Western Blot

    Diabetes and/or hyperglycemia via attenuation of the EGFR/erbB2 signaling and through subsequent modulation of downstream effectors such as ERK1/2, p38 MAPK or AKT/FOXO can lead to cardiac dysfunction. The effects of diabetes on EGFR/erbB2 pathway are exacerbated by blockade of this pathway by AG1478 or AG825 which leads to worsening cardiac recovery from I/R. However, the inhibitory effects of diabetes on EGFR/ErbB2 pathway may be opposed by administering EGF that also leads to improved cardiac function. The Angiotensin II (Ang II)/AT 1 receptor pathway can also activate EGFR/erbB2 pathway that can be blocked by Losartan. Co-administration of EGF with Losartan attenuates losartan-mediated EGFR blockade and improves cardiac function in diabetes beyond that attained by either drug alone.
    Figure Legend Snippet: Diabetes and/or hyperglycemia via attenuation of the EGFR/erbB2 signaling and through subsequent modulation of downstream effectors such as ERK1/2, p38 MAPK or AKT/FOXO can lead to cardiac dysfunction. The effects of diabetes on EGFR/erbB2 pathway are exacerbated by blockade of this pathway by AG1478 or AG825 which leads to worsening cardiac recovery from I/R. However, the inhibitory effects of diabetes on EGFR/ErbB2 pathway may be opposed by administering EGF that also leads to improved cardiac function. The Angiotensin II (Ang II)/AT 1 receptor pathway can also activate EGFR/erbB2 pathway that can be blocked by Losartan. Co-administration of EGF with Losartan attenuates losartan-mediated EGFR blockade and improves cardiac function in diabetes beyond that attained by either drug alone.

    Techniques Used:

    phospho her2 tyr1248 rabbit polyclonal antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho her2 tyr1248 rabbit polyclonal antibody
    SHP-1 binds to EGFR and HER-2 protein. Notes: ( A , B ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. EGFR and HER-2 proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( C , D ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. Phosphorylated-EGFR (pEGFR) and phosphorylated-HER-2 (pHER-2) proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( E , F ) The binding between SHP-1 and HER-3 or HER-4 was also evaluated through immunoprecipitation. The whole gel is shown, and no specific band of HER-3 or HER-4 was detected. ( G ) Bimolecular fluorescence complementation (BiFC) was performed to confirm the binding activity between SHP-1 and EGFR, HER-2, HER-3, and HER-4. BiFC signals were generated by interaction of the GFP fluorophore components based on proximity. + and − indicate receiving and withholding of the treatment, respectively. Abbreviations: EGFR, epidermal growth factor receptor; HER-2, human epidermal receptor; NC, nonsense control; WT, wild-type; IP, immunoprecipitation.
    Phospho Her2 Tyr1248 Rabbit Polyclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Protein tyrosine phosphatase SHP-1 sensitizes EGFR/HER-2 positive breast cancer cells to trastuzumab through modulating phosphorylation of EGFR and HER-2"

    Article Title: Protein tyrosine phosphatase SHP-1 sensitizes EGFR/HER-2 positive breast cancer cells to trastuzumab through modulating phosphorylation of EGFR and HER-2

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S82225

    SHP-1 binds to EGFR and HER-2 protein. Notes: ( A , B ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. EGFR and HER-2 proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( C , D ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. Phosphorylated-EGFR (pEGFR) and phosphorylated-HER-2 (pHER-2) proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( E , F ) The binding between SHP-1 and HER-3 or HER-4 was also evaluated through immunoprecipitation. The whole gel is shown, and no specific band of HER-3 or HER-4 was detected. ( G ) Bimolecular fluorescence complementation (BiFC) was performed to confirm the binding activity between SHP-1 and EGFR, HER-2, HER-3, and HER-4. BiFC signals were generated by interaction of the GFP fluorophore components based on proximity. + and − indicate receiving and withholding of the treatment, respectively. Abbreviations: EGFR, epidermal growth factor receptor; HER-2, human epidermal receptor; NC, nonsense control; WT, wild-type; IP, immunoprecipitation.
    Figure Legend Snippet: SHP-1 binds to EGFR and HER-2 protein. Notes: ( A , B ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. EGFR and HER-2 proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( C , D ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. Phosphorylated-EGFR (pEGFR) and phosphorylated-HER-2 (pHER-2) proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( E , F ) The binding between SHP-1 and HER-3 or HER-4 was also evaluated through immunoprecipitation. The whole gel is shown, and no specific band of HER-3 or HER-4 was detected. ( G ) Bimolecular fluorescence complementation (BiFC) was performed to confirm the binding activity between SHP-1 and EGFR, HER-2, HER-3, and HER-4. BiFC signals were generated by interaction of the GFP fluorophore components based on proximity. + and − indicate receiving and withholding of the treatment, respectively. Abbreviations: EGFR, epidermal growth factor receptor; HER-2, human epidermal receptor; NC, nonsense control; WT, wild-type; IP, immunoprecipitation.

    Techniques Used: Transfection, Plasmid Preparation, Immunoprecipitation, Western Blot, Binding Assay, Fluorescence, Activity Assay, Generated

    primary anti her2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc primary anti her2
    Primary Anti Her2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc p erbb2 tyr1248
    Primer sequences used for the qPCR analysis
    P Erbb2 Tyr1248, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    A–B, MdOS blocks <t>HER-2</t> phosphorylation and its downstream signal transduction in SK-OV-3 cells (A) and NIH-3T3/neu cells (B). C–D, Inhibition of EGF-stimulated EGFR phosphorylation and signal transduction by MdOS in A431 (C) and MDA-MB-468 (D) cells. Serum-starved cells were incubated with indicated concentrations of MdOS for 6 h at 37°C, EGF (50 ng/ml) was added to the cultures during the last 15-min treatment. E–F, Inhibition of VEGF-stimulated VEGFR2 phosphorylation and signal transduction by MdOS. HMEC (E) and NIH-3T3/flk-1 (F) cells were starved, then incubated with indicated concentrations of MdOS for 6 h at 37°C, VEGF (50 ng/ml) was added to the cultures during the last 15-min treatment. Protein samples were separated by SDS-PAGE and probed using the indicated antibodies. Representative data are shown.
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    a ) Representative Western blots showing levels of phosphorylated <t>erbB2</t> at Y877, Y1248, Y1248-a (which represents detection of Y1248 using an alternative antibody (p- erbB2-Antibody <t>(Tyr1248)/EGFR</t> (Tyr1173)) and Y12221/2 as well as total erbB2 (t-erbB2) and actin as a control protein in non-diabetic control hearts (C), diabetic hearts (D) and diabetic hearts chronically treated with AG825 (+AG825). b ) quantification of erbB2 expression relative to actin and c–f ) quantification of erbB2 phosphorylation at the stated tyrosine site relative to total erbB2 expression for all the groups studied by densitometry. N=4; * significantly different from control (p<0.05); ** significantly different from diabetes (p<0.05).
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    SHP-1 binds to EGFR and HER-2 protein. Notes: ( A , B ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. EGFR and HER-2 proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( C , D ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. Phosphorylated-EGFR (pEGFR) and phosphorylated-HER-2 (pHER-2) proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( E , F ) The binding between SHP-1 and HER-3 or HER-4 was also evaluated through immunoprecipitation. The whole gel is shown, and no specific band of HER-3 or HER-4 was detected. ( G ) Bimolecular fluorescence complementation (BiFC) was performed to confirm the binding activity between SHP-1 and EGFR, HER-2, HER-3, and HER-4. BiFC signals were generated by interaction of the GFP fluorophore components based on proximity. + and − indicate receiving and withholding of the treatment, respectively. Abbreviations: EGFR, epidermal growth factor receptor; HER-2, human epidermal receptor; NC, nonsense control; WT, wild-type; IP, immunoprecipitation.
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    SHP-1 binds to EGFR and HER-2 protein. Notes: ( A , B ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. EGFR and HER-2 proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( C , D ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. Phosphorylated-EGFR (pEGFR) and phosphorylated-HER-2 (pHER-2) proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( E , F ) The binding between SHP-1 and HER-3 or HER-4 was also evaluated through immunoprecipitation. The whole gel is shown, and no specific band of HER-3 or HER-4 was detected. ( G ) Bimolecular fluorescence complementation (BiFC) was performed to confirm the binding activity between SHP-1 and EGFR, HER-2, HER-3, and HER-4. BiFC signals were generated by interaction of the GFP fluorophore components based on proximity. + and − indicate receiving and withholding of the treatment, respectively. Abbreviations: EGFR, epidermal growth factor receptor; HER-2, human epidermal receptor; NC, nonsense control; WT, wild-type; IP, immunoprecipitation.
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    Image Search Results


    Primer sequences used for the qPCR analysis

    Journal: Drug Design, Development and Therapy

    Article Title: Dysregulation of Neuregulin-1/ErbB signaling in the hippocampus of rats after administration of doxorubicin

    doi: 10.2147/DDDT.S151511

    Figure Lengend Snippet: Primer sequences used for the qPCR analysis

    Article Snippet: The following antibodies and concentrations were used over night at 4°C; NRG1 (Santa Cruz Biotechnology Inc., Dallas, TX, USA, sc-28916; 1:400), ErbB4 (Santa Cruz Biotechnology Inc., sc-283; 1:500), ErbB2 (Cell Signaling, Danvers, MA, USA, 2165; 1:1,500), p-ErbB4 (Tyr1056) (Santa Cruz Biotechnology Inc., sc33040; 1:300), p-ErbB2 (Tyr1248) (Cell Signaling, 2247; 1:1,500), p-Akt (Ser473) (Cell Signaling, 4060; 1:3,000), p-ERK (Thr202/Tyr204) (Cell Signaling, 4695; 1:2,000), caspase-3 (Abcam, Cambridge, UK, ab4051; 1:500), and β-actin (Proteintech, Rosemont, IL, USA, 66009-1-Ig; 1:4,000).

    Techniques: Amplification

    Effect of Dox on gene expression of ErbB2, ErbB4, and the ratio of pErbB4/ErbB4 and pErbB2/ErbB2 in the hippocampus. Notes: ErbB4 mRNA expression ( A ); pErbB4/ErbB4 ratio ( B ); ErbB2 mRNA expression ( C ); and pErbB2/ErbB2 ratio ( D ). Data are expressed as mean ± SEM (n=6–7). * p <0.05 and ** p <0.01 compared to the control group. Abbreviations: Dox, doxorubicin; DoxS, doxorubicin administration for short time; DoxL, doxorubicin administration for long time; SEM, standard error of the mean.

    Journal: Drug Design, Development and Therapy

    Article Title: Dysregulation of Neuregulin-1/ErbB signaling in the hippocampus of rats after administration of doxorubicin

    doi: 10.2147/DDDT.S151511

    Figure Lengend Snippet: Effect of Dox on gene expression of ErbB2, ErbB4, and the ratio of pErbB4/ErbB4 and pErbB2/ErbB2 in the hippocampus. Notes: ErbB4 mRNA expression ( A ); pErbB4/ErbB4 ratio ( B ); ErbB2 mRNA expression ( C ); and pErbB2/ErbB2 ratio ( D ). Data are expressed as mean ± SEM (n=6–7). * p <0.05 and ** p <0.01 compared to the control group. Abbreviations: Dox, doxorubicin; DoxS, doxorubicin administration for short time; DoxL, doxorubicin administration for long time; SEM, standard error of the mean.

    Article Snippet: The following antibodies and concentrations were used over night at 4°C; NRG1 (Santa Cruz Biotechnology Inc., Dallas, TX, USA, sc-28916; 1:400), ErbB4 (Santa Cruz Biotechnology Inc., sc-283; 1:500), ErbB2 (Cell Signaling, Danvers, MA, USA, 2165; 1:1,500), p-ErbB4 (Tyr1056) (Santa Cruz Biotechnology Inc., sc33040; 1:300), p-ErbB2 (Tyr1248) (Cell Signaling, 2247; 1:1,500), p-Akt (Ser473) (Cell Signaling, 4060; 1:3,000), p-ERK (Thr202/Tyr204) (Cell Signaling, 4695; 1:2,000), caspase-3 (Abcam, Cambridge, UK, ab4051; 1:500), and β-actin (Proteintech, Rosemont, IL, USA, 66009-1-Ig; 1:4,000).

    Techniques: Expressing

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: KRAS G12C -independent feedback activation of wild-type RAS constrains KRAS G12C inhibitor efficacy

    doi: 10.1016/j.celrep.2022.110993

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Rabbit monoclonal Anti-phospho-HER2 Y1248 , Cell Signaling Technologies , Cat# 2247S.

    Techniques: Derivative Assay, Recombinant, In Vitro, In Vivo, Software

    A–B, MdOS blocks HER-2 phosphorylation and its downstream signal transduction in SK-OV-3 cells (A) and NIH-3T3/neu cells (B). C–D, Inhibition of EGF-stimulated EGFR phosphorylation and signal transduction by MdOS in A431 (C) and MDA-MB-468 (D) cells. Serum-starved cells were incubated with indicated concentrations of MdOS for 6 h at 37°C, EGF (50 ng/ml) was added to the cultures during the last 15-min treatment. E–F, Inhibition of VEGF-stimulated VEGFR2 phosphorylation and signal transduction by MdOS. HMEC (E) and NIH-3T3/flk-1 (F) cells were starved, then incubated with indicated concentrations of MdOS for 6 h at 37°C, VEGF (50 ng/ml) was added to the cultures during the last 15-min treatment. Protein samples were separated by SDS-PAGE and probed using the indicated antibodies. Representative data are shown.

    Journal: PLoS ONE

    Article Title: The Marine-Derived Oligosaccharide Sulfate (MdOS), a Novel Multiple Tyrosine Kinase Inhibitor, Combats Tumor Angiogenesis both In Vitro and In Vivo

    doi: 10.1371/journal.pone.0003774

    Figure Lengend Snippet: A–B, MdOS blocks HER-2 phosphorylation and its downstream signal transduction in SK-OV-3 cells (A) and NIH-3T3/neu cells (B). C–D, Inhibition of EGF-stimulated EGFR phosphorylation and signal transduction by MdOS in A431 (C) and MDA-MB-468 (D) cells. Serum-starved cells were incubated with indicated concentrations of MdOS for 6 h at 37°C, EGF (50 ng/ml) was added to the cultures during the last 15-min treatment. E–F, Inhibition of VEGF-stimulated VEGFR2 phosphorylation and signal transduction by MdOS. HMEC (E) and NIH-3T3/flk-1 (F) cells were starved, then incubated with indicated concentrations of MdOS for 6 h at 37°C, VEGF (50 ng/ml) was added to the cultures during the last 15-min treatment. Protein samples were separated by SDS-PAGE and probed using the indicated antibodies. Representative data are shown.

    Article Snippet: Western blot analyses were subsequently performed as previous described , with the antibodies against phospho-EGFR (Tyr1068) (#2234), phospho-HER-2 (Tyr1248) (#2247), phospho-VEGFR2 (Tyr996) (#2474), VEGFR2 (#2472), phospho-ERK1/2 (#9101), ERK1/2 (#9102), phosphor-AKT (Ser473) (#9271) and AKT (#9272) (Cell Signaling Technology, Beverly, MA, USA), EGFR (sc-03), HER-2(sc-284) and actin (sc-8432) (Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: Transduction, Inhibition, Incubation, SDS Page

    VEGFR2 was injected at the concentrations of 2.56, 0.64, 0.32, 0.16 and 0.08 µM (from top to bottom); HER-2 was injected at the concentrations of 0.53, 0.26, 0.13, 0.07 and 0.03 µM (from top to bottom); EGFR was injected at the concentrations of 6, 3, 1.5, 0.75, 0.38, 0.19 and 0.09 µM (from top to bottom); 6×his-tag was injected at the concentrations of 100, 10, 1, 0.1, 0.01 µM. Sensorgram responses at equilibrium were plotted against each concentration of compounds and the equilibrium dissociation constant (K D ) of the binding system was calculated using BIAeval software 3.1.

    Journal: PLoS ONE

    Article Title: The Marine-Derived Oligosaccharide Sulfate (MdOS), a Novel Multiple Tyrosine Kinase Inhibitor, Combats Tumor Angiogenesis both In Vitro and In Vivo

    doi: 10.1371/journal.pone.0003774

    Figure Lengend Snippet: VEGFR2 was injected at the concentrations of 2.56, 0.64, 0.32, 0.16 and 0.08 µM (from top to bottom); HER-2 was injected at the concentrations of 0.53, 0.26, 0.13, 0.07 and 0.03 µM (from top to bottom); EGFR was injected at the concentrations of 6, 3, 1.5, 0.75, 0.38, 0.19 and 0.09 µM (from top to bottom); 6×his-tag was injected at the concentrations of 100, 10, 1, 0.1, 0.01 µM. Sensorgram responses at equilibrium were plotted against each concentration of compounds and the equilibrium dissociation constant (K D ) of the binding system was calculated using BIAeval software 3.1.

    Article Snippet: Western blot analyses were subsequently performed as previous described , with the antibodies against phospho-EGFR (Tyr1068) (#2234), phospho-HER-2 (Tyr1248) (#2247), phospho-VEGFR2 (Tyr996) (#2474), VEGFR2 (#2472), phospho-ERK1/2 (#9101), ERK1/2 (#9102), phosphor-AKT (Ser473) (#9271) and AKT (#9272) (Cell Signaling Technology, Beverly, MA, USA), EGFR (sc-03), HER-2(sc-284) and actin (sc-8432) (Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: Injection, Concentration Assay, Binding Assay, Software

    VEGFR2 (A) and HER-2 (B) and EGFR (C) kinase assays were performed as described in in the presence of varying concentrations of ATP. Initial reaction velocity was expressed as the phosphorylation of poly(Glu, Tyr) 4∶1 substrate. All x, y data sets were multiplied by 100 for purposes of graphical presentation.

    Journal: PLoS ONE

    Article Title: The Marine-Derived Oligosaccharide Sulfate (MdOS), a Novel Multiple Tyrosine Kinase Inhibitor, Combats Tumor Angiogenesis both In Vitro and In Vivo

    doi: 10.1371/journal.pone.0003774

    Figure Lengend Snippet: VEGFR2 (A) and HER-2 (B) and EGFR (C) kinase assays were performed as described in in the presence of varying concentrations of ATP. Initial reaction velocity was expressed as the phosphorylation of poly(Glu, Tyr) 4∶1 substrate. All x, y data sets were multiplied by 100 for purposes of graphical presentation.

    Article Snippet: Western blot analyses were subsequently performed as previous described , with the antibodies against phospho-EGFR (Tyr1068) (#2234), phospho-HER-2 (Tyr1248) (#2247), phospho-VEGFR2 (Tyr996) (#2474), VEGFR2 (#2472), phospho-ERK1/2 (#9101), ERK1/2 (#9102), phosphor-AKT (Ser473) (#9271) and AKT (#9272) (Cell Signaling Technology, Beverly, MA, USA), EGFR (sc-03), HER-2(sc-284) and actin (sc-8432) (Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques:

    a ) Representative Western blots showing levels of phosphorylated erbB2 at Y877, Y1248, Y1248-a (which represents detection of Y1248 using an alternative antibody (p- erbB2-Antibody (Tyr1248)/EGFR (Tyr1173)) and Y12221/2 as well as total erbB2 (t-erbB2) and actin as a control protein in non-diabetic control hearts (C), diabetic hearts (D) and diabetic hearts chronically treated with AG825 (+AG825). b ) quantification of erbB2 expression relative to actin and c–f ) quantification of erbB2 phosphorylation at the stated tyrosine site relative to total erbB2 expression for all the groups studied by densitometry. N=4; * significantly different from control (p<0.05); ** significantly different from diabetes (p<0.05).

    Journal: PLoS ONE

    Article Title: Activation of EGFR/ERBB2 via Pathways Involving ERK1/2, P38 MAPK, AKT and FOXO Enhances Recovery of Diabetic Hearts from Ischemia-Reperfusion Injury

    doi: 10.1371/journal.pone.0039066

    Figure Lengend Snippet: a ) Representative Western blots showing levels of phosphorylated erbB2 at Y877, Y1248, Y1248-a (which represents detection of Y1248 using an alternative antibody (p- erbB2-Antibody (Tyr1248)/EGFR (Tyr1173)) and Y12221/2 as well as total erbB2 (t-erbB2) and actin as a control protein in non-diabetic control hearts (C), diabetic hearts (D) and diabetic hearts chronically treated with AG825 (+AG825). b ) quantification of erbB2 expression relative to actin and c–f ) quantification of erbB2 phosphorylation at the stated tyrosine site relative to total erbB2 expression for all the groups studied by densitometry. N=4; * significantly different from control (p<0.05); ** significantly different from diabetes (p<0.05).

    Article Snippet: The following antibodies from Cell Signaling (USA) were used in this study: t-EGFR-Antibody (rabbit) Cat. No. 2232, p-EGFR-Antibody (Tyr992) (rabbit) Cat. No. 2235, p-EGFR-Antibody (Tyr1068) (rabbit) Cat. No. 2234, p-EGFR-Antibody (Tyr1086) (rabbit) Cat. No. 2220, p-EGFR-Antibody (Tyr1148) (rabbit) Cat. No. 4404, t-Her2/ErbB2-Antibody (29D8) (rabbit) Cat. No. 2165, p-Her2/ErbB2-Antibody (Tyr877) (rabbit) Cat. No. 2241, p-Her2/ErbB2-Antibody (Tyr1248) (rabbit) Cat. No. 2247, p-Her2/ErbB2-Antibody (Tyr1248)/EGFR(Tyr1173) (rabbit) Cat. No. 2244, p-Her2/ErbB2-Antibody (Tyr1221/1222) (rabbit) Cat. No. 2243, p-Foxo3a (Ser 253) Cat. No. 9466, p-ERK1/2 (p44/42 MAP Kinase, Thr202/Tyr204) Antibody (rabbit) Cat. No. 9101.

    Techniques: Western Blot, Expressing

    Panel a) are represenatative Western Blots following immunoprecipitations (IP) with either total-EGFR or total-erbB2 antibody and subsequent immunoblotting (IB) with both antibodies individually. Panel b) represents the mean ratio of erbB2/EGFR dimers as assessed by densitometry for non-diabetic control hearts (C), diabetic hearts (D) and diabetic hearts chronically treated with AG1478 (+AG1478) or AG825 (+AG825). N=4; * significantly different from control (p<0.05); ** significantly different from diabetes (p<0.05).

    Journal: PLoS ONE

    Article Title: Activation of EGFR/ERBB2 via Pathways Involving ERK1/2, P38 MAPK, AKT and FOXO Enhances Recovery of Diabetic Hearts from Ischemia-Reperfusion Injury

    doi: 10.1371/journal.pone.0039066

    Figure Lengend Snippet: Panel a) are represenatative Western Blots following immunoprecipitations (IP) with either total-EGFR or total-erbB2 antibody and subsequent immunoblotting (IB) with both antibodies individually. Panel b) represents the mean ratio of erbB2/EGFR dimers as assessed by densitometry for non-diabetic control hearts (C), diabetic hearts (D) and diabetic hearts chronically treated with AG1478 (+AG1478) or AG825 (+AG825). N=4; * significantly different from control (p<0.05); ** significantly different from diabetes (p<0.05).

    Article Snippet: The following antibodies from Cell Signaling (USA) were used in this study: t-EGFR-Antibody (rabbit) Cat. No. 2232, p-EGFR-Antibody (Tyr992) (rabbit) Cat. No. 2235, p-EGFR-Antibody (Tyr1068) (rabbit) Cat. No. 2234, p-EGFR-Antibody (Tyr1086) (rabbit) Cat. No. 2220, p-EGFR-Antibody (Tyr1148) (rabbit) Cat. No. 4404, t-Her2/ErbB2-Antibody (29D8) (rabbit) Cat. No. 2165, p-Her2/ErbB2-Antibody (Tyr877) (rabbit) Cat. No. 2241, p-Her2/ErbB2-Antibody (Tyr1248) (rabbit) Cat. No. 2247, p-Her2/ErbB2-Antibody (Tyr1248)/EGFR(Tyr1173) (rabbit) Cat. No. 2244, p-Her2/ErbB2-Antibody (Tyr1221/1222) (rabbit) Cat. No. 2243, p-Foxo3a (Ser 253) Cat. No. 9466, p-ERK1/2 (p44/42 MAP Kinase, Thr202/Tyr204) Antibody (rabbit) Cat. No. 9101.

    Techniques: Western Blot

    Representative Western blots to show ischemia-induced changes in phosphorylation of EGFR and erbB2 receptors and downstream signaling molecules for a) normal (non-diabetic) control hearts (C) and those subjected to 40 min ischemia (CI) and b) diabetic hearts (D) and those subjected to 40 min ischemia (DI).

    Journal: PLoS ONE

    Article Title: Activation of EGFR/ERBB2 via Pathways Involving ERK1/2, P38 MAPK, AKT and FOXO Enhances Recovery of Diabetic Hearts from Ischemia-Reperfusion Injury

    doi: 10.1371/journal.pone.0039066

    Figure Lengend Snippet: Representative Western blots to show ischemia-induced changes in phosphorylation of EGFR and erbB2 receptors and downstream signaling molecules for a) normal (non-diabetic) control hearts (C) and those subjected to 40 min ischemia (CI) and b) diabetic hearts (D) and those subjected to 40 min ischemia (DI).

    Article Snippet: The following antibodies from Cell Signaling (USA) were used in this study: t-EGFR-Antibody (rabbit) Cat. No. 2232, p-EGFR-Antibody (Tyr992) (rabbit) Cat. No. 2235, p-EGFR-Antibody (Tyr1068) (rabbit) Cat. No. 2234, p-EGFR-Antibody (Tyr1086) (rabbit) Cat. No. 2220, p-EGFR-Antibody (Tyr1148) (rabbit) Cat. No. 4404, t-Her2/ErbB2-Antibody (29D8) (rabbit) Cat. No. 2165, p-Her2/ErbB2-Antibody (Tyr877) (rabbit) Cat. No. 2241, p-Her2/ErbB2-Antibody (Tyr1248) (rabbit) Cat. No. 2247, p-Her2/ErbB2-Antibody (Tyr1248)/EGFR(Tyr1173) (rabbit) Cat. No. 2244, p-Her2/ErbB2-Antibody (Tyr1221/1222) (rabbit) Cat. No. 2243, p-Foxo3a (Ser 253) Cat. No. 9466, p-ERK1/2 (p44/42 MAP Kinase, Thr202/Tyr204) Antibody (rabbit) Cat. No. 9101.

    Techniques: Western Blot

    Diabetes and/or hyperglycemia via attenuation of the EGFR/erbB2 signaling and through subsequent modulation of downstream effectors such as ERK1/2, p38 MAPK or AKT/FOXO can lead to cardiac dysfunction. The effects of diabetes on EGFR/erbB2 pathway are exacerbated by blockade of this pathway by AG1478 or AG825 which leads to worsening cardiac recovery from I/R. However, the inhibitory effects of diabetes on EGFR/ErbB2 pathway may be opposed by administering EGF that also leads to improved cardiac function. The Angiotensin II (Ang II)/AT 1 receptor pathway can also activate EGFR/erbB2 pathway that can be blocked by Losartan. Co-administration of EGF with Losartan attenuates losartan-mediated EGFR blockade and improves cardiac function in diabetes beyond that attained by either drug alone.

    Journal: PLoS ONE

    Article Title: Activation of EGFR/ERBB2 via Pathways Involving ERK1/2, P38 MAPK, AKT and FOXO Enhances Recovery of Diabetic Hearts from Ischemia-Reperfusion Injury

    doi: 10.1371/journal.pone.0039066

    Figure Lengend Snippet: Diabetes and/or hyperglycemia via attenuation of the EGFR/erbB2 signaling and through subsequent modulation of downstream effectors such as ERK1/2, p38 MAPK or AKT/FOXO can lead to cardiac dysfunction. The effects of diabetes on EGFR/erbB2 pathway are exacerbated by blockade of this pathway by AG1478 or AG825 which leads to worsening cardiac recovery from I/R. However, the inhibitory effects of diabetes on EGFR/ErbB2 pathway may be opposed by administering EGF that also leads to improved cardiac function. The Angiotensin II (Ang II)/AT 1 receptor pathway can also activate EGFR/erbB2 pathway that can be blocked by Losartan. Co-administration of EGF with Losartan attenuates losartan-mediated EGFR blockade and improves cardiac function in diabetes beyond that attained by either drug alone.

    Article Snippet: The following antibodies from Cell Signaling (USA) were used in this study: t-EGFR-Antibody (rabbit) Cat. No. 2232, p-EGFR-Antibody (Tyr992) (rabbit) Cat. No. 2235, p-EGFR-Antibody (Tyr1068) (rabbit) Cat. No. 2234, p-EGFR-Antibody (Tyr1086) (rabbit) Cat. No. 2220, p-EGFR-Antibody (Tyr1148) (rabbit) Cat. No. 4404, t-Her2/ErbB2-Antibody (29D8) (rabbit) Cat. No. 2165, p-Her2/ErbB2-Antibody (Tyr877) (rabbit) Cat. No. 2241, p-Her2/ErbB2-Antibody (Tyr1248) (rabbit) Cat. No. 2247, p-Her2/ErbB2-Antibody (Tyr1248)/EGFR(Tyr1173) (rabbit) Cat. No. 2244, p-Her2/ErbB2-Antibody (Tyr1221/1222) (rabbit) Cat. No. 2243, p-Foxo3a (Ser 253) Cat. No. 9466, p-ERK1/2 (p44/42 MAP Kinase, Thr202/Tyr204) Antibody (rabbit) Cat. No. 9101.

    Techniques:

    SHP-1 binds to EGFR and HER-2 protein. Notes: ( A , B ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. EGFR and HER-2 proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( C , D ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. Phosphorylated-EGFR (pEGFR) and phosphorylated-HER-2 (pHER-2) proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( E , F ) The binding between SHP-1 and HER-3 or HER-4 was also evaluated through immunoprecipitation. The whole gel is shown, and no specific band of HER-3 or HER-4 was detected. ( G ) Bimolecular fluorescence complementation (BiFC) was performed to confirm the binding activity between SHP-1 and EGFR, HER-2, HER-3, and HER-4. BiFC signals were generated by interaction of the GFP fluorophore components based on proximity. + and − indicate receiving and withholding of the treatment, respectively. Abbreviations: EGFR, epidermal growth factor receptor; HER-2, human epidermal receptor; NC, nonsense control; WT, wild-type; IP, immunoprecipitation.

    Journal: OncoTargets and therapy

    Article Title: Protein tyrosine phosphatase SHP-1 sensitizes EGFR/HER-2 positive breast cancer cells to trastuzumab through modulating phosphorylation of EGFR and HER-2

    doi: 10.2147/OTT.S82225

    Figure Lengend Snippet: SHP-1 binds to EGFR and HER-2 protein. Notes: ( A , B ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. EGFR and HER-2 proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( C , D ) SKBR3 cells were transfected with control (NC), SHP-1 WT, or SHP-1 MT vector for 48 hours. Phosphorylated-EGFR (pEGFR) and phosphorylated-HER-2 (pHER-2) proteins were pulled down by immunoprecipitation and the protein level was detected by immunoblotting. ( E , F ) The binding between SHP-1 and HER-3 or HER-4 was also evaluated through immunoprecipitation. The whole gel is shown, and no specific band of HER-3 or HER-4 was detected. ( G ) Bimolecular fluorescence complementation (BiFC) was performed to confirm the binding activity between SHP-1 and EGFR, HER-2, HER-3, and HER-4. BiFC signals were generated by interaction of the GFP fluorophore components based on proximity. + and − indicate receiving and withholding of the treatment, respectively. Abbreviations: EGFR, epidermal growth factor receptor; HER-2, human epidermal receptor; NC, nonsense control; WT, wild-type; IP, immunoprecipitation.

    Article Snippet: All antibodies were purchased from Cell Signaling Technologies: phospho-HER2 (Tyr1248) rabbit polyclonal antibody (#2247), HER2 rabbit polyclonal antibody (#2242), phospho-EGFR (Tyr1173) rabbit monoclonal antibody (#4407), EGFR rabbit monoclonal antibody (#4405), phospho-Erk1/2 (Thr202/Tyr204) rabbit monoclonal antibody (#4370), Erk1/2 rabbit monoclonal antibody (#4695), phospho-Stat3 (Tyr705) rabbit monoclonal antibody (#9145), Stat3 rabbit monoclonal antibody (#4904), phospho-Akt (Ser473) rabbit monoclonal antibody (#4060), and Akt (pan) rabbit monoclonal antibody (#4685).

    Techniques: Transfection, Plasmid Preparation, Immunoprecipitation, Western Blot, Binding Assay, Fluorescence, Activity Assay, Generated