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pbabe puro egfr wt  (Addgene inc)


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    Addgene inc pbabe puro egfr wt
    Pbabe Puro Egfr Wt, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 10 article reviews
    pbabe puro egfr wt - by Bioz Stars, 2026-02
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    A, Immunofluorescent images of MCF10A cells from in situ proximity ligation assays using primary antibodies against EGFR and HER2 to assess co-localization. Cells are parental or overexpress exogenous cDNAs of EGFR WT, EGFR N361A, EGFR <t>L858R,</t> or a single construct containing two mutations EGFR N361A/L858R. Scale bar represents 50 μm. B, Representative quantification of mean pixel intensity of cellular regions defined by rhodamine Actin stain, including five replicate images per condition. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, n = 5.
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    A, Diagram of domain architecture of <t>EGFR,</t> showing heterodimerization between EGFR and a binding partner such as Her2. Yellow star indicates approximate site of the common oncogenic L858 mutation, red star indicates approximate site of the N361 glycosylation site, green indicates ligands such as EGF or AREG that bind to EGFR to induce dimerization. B, Percentage of fluorescent cells from flow cytometry of MCF10A cells that were labeled with anti-EGFR antibody conjugated to Alexa-Fluor 488, or empty <t>puro-IRES-eGFP</t> vector as the stably fluorescent positive control which is not labeled by fluorescent antibody. Cells used were parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR wild-type (WT) or EGFR N361A. C, Representative immunofluorescent microscopy images of parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR wild-type (WT) or EGFR N361A, stained for EGFR, vimentin, ATP1A1, or DAPI.
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    A, Diagram of domain architecture of <t>EGFR,</t> showing heterodimerization between EGFR and a binding partner such as Her2. Yellow star indicates approximate site of the common oncogenic L858 mutation, red star indicates approximate site of the N361 glycosylation site, green indicates ligands such as EGF or AREG that bind to EGFR to induce dimerization. B, Percentage of fluorescent cells from flow cytometry of MCF10A cells that were labeled with anti-EGFR antibody conjugated to Alexa-Fluor 488, or empty <t>puro-IRES-eGFP</t> vector as the stably fluorescent positive control which is not labeled by fluorescent antibody. Cells used were parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR wild-type (WT) or EGFR N361A. C, Representative immunofluorescent microscopy images of parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR wild-type (WT) or EGFR N361A, stained for EGFR, vimentin, ATP1A1, or DAPI.
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    A, Diagram of domain architecture of <t>EGFR,</t> showing heterodimerization between EGFR and a binding partner such as Her2. Yellow star indicates approximate site of the common oncogenic L858 mutation, red star indicates approximate site of the N361 glycosylation site, green indicates ligands such as EGF or AREG that bind to EGFR to induce dimerization. B, Percentage of fluorescent cells from flow cytometry of MCF10A cells that were labeled with anti-EGFR antibody conjugated to Alexa-Fluor 488, or empty <t>puro-IRES-eGFP</t> vector as the stably fluorescent positive control which is not labeled by fluorescent antibody. Cells used were parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR wild-type (WT) or EGFR N361A. C, Representative immunofluorescent microscopy images of parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR wild-type (WT) or EGFR N361A, stained for EGFR, vimentin, ATP1A1, or DAPI.
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    A, Immunofluorescent images of MCF10A cells from in situ proximity ligation assays using primary antibodies against EGFR and HER2 to assess co-localization. Cells are parental or overexpress exogenous cDNAs of EGFR WT, EGFR N361A, EGFR L858R, or a single construct containing two mutations EGFR N361A/L858R. Scale bar represents 50 μm. B, Representative quantification of mean pixel intensity of cellular regions defined by rhodamine Actin stain, including five replicate images per condition. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, n = 5.

    Journal: bioRxiv

    Article Title: Effects of N361 Glycosylation on Epidermal Growth Factor Receptor Biological Function

    doi: 10.1101/2024.07.12.603279

    Figure Lengend Snippet: A, Immunofluorescent images of MCF10A cells from in situ proximity ligation assays using primary antibodies against EGFR and HER2 to assess co-localization. Cells are parental or overexpress exogenous cDNAs of EGFR WT, EGFR N361A, EGFR L858R, or a single construct containing two mutations EGFR N361A/L858R. Scale bar represents 50 μm. B, Representative quantification of mean pixel intensity of cellular regions defined by rhodamine Actin stain, including five replicate images per condition. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, n = 5.

    Article Snippet: Introduction of N361A by site-directed mutagenesis on the pBABE puro EGFR wild-type (EGFR WT, RRID:Addgene_11011) and pBABE puro EGFR L858R (RRID:Addgene_11012) plasmids was performed by Azenta. pBABE puro IRES-eGFP (RRID:Addgene_14430).

    Techniques: In Situ, Ligation, Construct, Staining

    A, Diagram of domain architecture of EGFR, showing heterodimerization between EGFR and a binding partner such as Her2. Yellow star indicates approximate site of the common oncogenic L858 mutation, red star indicates approximate site of the N361 glycosylation site, green indicates ligands such as EGF or AREG that bind to EGFR to induce dimerization. B, Percentage of fluorescent cells from flow cytometry of MCF10A cells that were labeled with anti-EGFR antibody conjugated to Alexa-Fluor 488, or empty puro-IRES-eGFP vector as the stably fluorescent positive control which is not labeled by fluorescent antibody. Cells used were parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR wild-type (WT) or EGFR N361A. C, Representative immunofluorescent microscopy images of parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR wild-type (WT) or EGFR N361A, stained for EGFR, vimentin, ATP1A1, or DAPI.

    Journal: bioRxiv

    Article Title: Effects of N361 Glycosylation on Epidermal Growth Factor Receptor Biological Function

    doi: 10.1101/2024.07.12.603279

    Figure Lengend Snippet: A, Diagram of domain architecture of EGFR, showing heterodimerization between EGFR and a binding partner such as Her2. Yellow star indicates approximate site of the common oncogenic L858 mutation, red star indicates approximate site of the N361 glycosylation site, green indicates ligands such as EGF or AREG that bind to EGFR to induce dimerization. B, Percentage of fluorescent cells from flow cytometry of MCF10A cells that were labeled with anti-EGFR antibody conjugated to Alexa-Fluor 488, or empty puro-IRES-eGFP vector as the stably fluorescent positive control which is not labeled by fluorescent antibody. Cells used were parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR wild-type (WT) or EGFR N361A. C, Representative immunofluorescent microscopy images of parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR wild-type (WT) or EGFR N361A, stained for EGFR, vimentin, ATP1A1, or DAPI.

    Article Snippet: Introduction of N361A by site-directed mutagenesis on the pBABE puro EGFR wild-type (EGFR WT, RRID:Addgene_11011) and pBABE puro EGFR L858R (RRID:Addgene_11012) plasmids was performed by Azenta. pBABE puro IRES-eGFP (RRID:Addgene_14430).

    Techniques: Binding Assay, Mutagenesis, Glycoproteomics, Flow Cytometry, Labeling, Plasmid Preparation, Stable Transfection, Positive Control, Microscopy, Staining

    A, Time course of relative viability of in parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR WT or EGFR N361A, measured by Cell-Titer Glo luciferase assay (CTG) in stimulated media with 20 ng/mL of EGF. B, Dose course of relative viability of MCF10A cells overexpressing the indicated constructs after stimulation by EGF for 72 hours, measured by CTG. C, Dose course of relative viability of parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR WT or EGFR N361A upon stimulation by AREG for 72 hours, both measured by CTG; ns = not significant, * p < 0.05; *** p < 0.001, n = 3; D, Immunoblots of whole cell lysates of parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR WT or EGFR N361A stimulated with an additional 20 ng/mL of EGF for 15 minutes.

    Journal: bioRxiv

    Article Title: Effects of N361 Glycosylation on Epidermal Growth Factor Receptor Biological Function

    doi: 10.1101/2024.07.12.603279

    Figure Lengend Snippet: A, Time course of relative viability of in parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR WT or EGFR N361A, measured by Cell-Titer Glo luciferase assay (CTG) in stimulated media with 20 ng/mL of EGF. B, Dose course of relative viability of MCF10A cells overexpressing the indicated constructs after stimulation by EGF for 72 hours, measured by CTG. C, Dose course of relative viability of parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR WT or EGFR N361A upon stimulation by AREG for 72 hours, both measured by CTG; ns = not significant, * p < 0.05; *** p < 0.001, n = 3; D, Immunoblots of whole cell lysates of parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR WT or EGFR N361A stimulated with an additional 20 ng/mL of EGF for 15 minutes.

    Article Snippet: Introduction of N361A by site-directed mutagenesis on the pBABE puro EGFR wild-type (EGFR WT, RRID:Addgene_11011) and pBABE puro EGFR L858R (RRID:Addgene_11012) plasmids was performed by Azenta. pBABE puro IRES-eGFP (RRID:Addgene_14430).

    Techniques: Luciferase, Construct, Western Blot

    A, Immunofluorescent images of MCF10A cells from in situ proximity ligation assays using primary antibodies against EGFR and HER2 to assess co-localization. Cells are parental or overexpress exogenous cDNAs of EGFR WT, EGFR N361A, EGFR L858R, or a single construct containing two mutations EGFR N361A/L858R. Scale bar represents 50 μm. B, Representative quantification of mean pixel intensity of cellular regions defined by rhodamine Actin stain, including five replicate images per condition. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, n = 5.

    Journal: bioRxiv

    Article Title: Effects of N361 Glycosylation on Epidermal Growth Factor Receptor Biological Function

    doi: 10.1101/2024.07.12.603279

    Figure Lengend Snippet: A, Immunofluorescent images of MCF10A cells from in situ proximity ligation assays using primary antibodies against EGFR and HER2 to assess co-localization. Cells are parental or overexpress exogenous cDNAs of EGFR WT, EGFR N361A, EGFR L858R, or a single construct containing two mutations EGFR N361A/L858R. Scale bar represents 50 μm. B, Representative quantification of mean pixel intensity of cellular regions defined by rhodamine Actin stain, including five replicate images per condition. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, n = 5.

    Article Snippet: Introduction of N361A by site-directed mutagenesis on the pBABE puro EGFR wild-type (EGFR WT, RRID:Addgene_11011) and pBABE puro EGFR L858R (RRID:Addgene_11012) plasmids was performed by Azenta. pBABE puro IRES-eGFP (RRID:Addgene_14430).

    Techniques: In Situ, Ligation, Construct, Staining

    A-B, Dose course of relative viability measured by CTG of parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR WT or EGFR N361A after 72 hours of treatment with either (A) necitumumab or (B) osimertinib. ns = not significant, * p < 0.05; *** p < 0.001, n = 3.

    Journal: bioRxiv

    Article Title: Effects of N361 Glycosylation on Epidermal Growth Factor Receptor Biological Function

    doi: 10.1101/2024.07.12.603279

    Figure Lengend Snippet: A-B, Dose course of relative viability measured by CTG of parental MCF10A cells or MCF10A cells overexpressing cDNAs of EGFR WT or EGFR N361A after 72 hours of treatment with either (A) necitumumab or (B) osimertinib. ns = not significant, * p < 0.05; *** p < 0.001, n = 3.

    Article Snippet: Introduction of N361A by site-directed mutagenesis on the pBABE puro EGFR wild-type (EGFR WT, RRID:Addgene_11011) and pBABE puro EGFR L858R (RRID:Addgene_11012) plasmids was performed by Azenta. pBABE puro IRES-eGFP (RRID:Addgene_14430).

    Techniques: