p egfr  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p egfr
    P Egfr, 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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    p egfr  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p egfr
    P Egfr, 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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    p egfr  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p egfr
    A). ST6GAL1 was stably knocked-down (KD) in cells with high endogenous ST6GAL1 expression (MiaPaCa-2, S2-013, S2-LM7AA, OVCAR-3, and OVCAR-5) using lentivirus encoding an shRNA sequence targeting ST6GAL1. As controls, cells were either transduced with lentivirus containing shRNA targeting GFP (shC) or with an empty vector (EV) construct. B). Cells with undetectable endogenous ST6GAL1 (SW48 and OV4) were stably transduced with ST6GAL1-encoding cDNA to overexpress (OE) the enzyme, or with an EV construct. All cell lines represent polyclonal populations. C). Cells with or without ST6GAL1 KD were treated with 100 ng/mL EGF for 15 minutes and immunoblotted <t>for</t> <t>p-EGFR</t> (pY1068) and total EGFR (t-EGFR). D). Cells with or without ST6GAL1 OE were treated with 100 ng/mL EGF for 15 minutes and immunoblotted for p-EGFR and t-EGFR.
    P Egfr, 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 "Sialylation of EGFR by ST6GAL1 induces receptor activation and modulates trafficking dynamics"

    Article Title: Sialylation of EGFR by ST6GAL1 induces receptor activation and modulates trafficking dynamics

    Journal: bioRxiv

    doi: 10.1101/2023.06.03.543566

    A). ST6GAL1 was stably knocked-down (KD) in cells with high endogenous ST6GAL1 expression (MiaPaCa-2, S2-013, S2-LM7AA, OVCAR-3, and OVCAR-5) using lentivirus encoding an shRNA sequence targeting ST6GAL1. As controls, cells were either transduced with lentivirus containing shRNA targeting GFP (shC) or with an empty vector (EV) construct. B). Cells with undetectable endogenous ST6GAL1 (SW48 and OV4) were stably transduced with ST6GAL1-encoding cDNA to overexpress (OE) the enzyme, or with an EV construct. All cell lines represent polyclonal populations. C). Cells with or without ST6GAL1 KD were treated with 100 ng/mL EGF for 15 minutes and immunoblotted for p-EGFR (pY1068) and total EGFR (t-EGFR). D). Cells with or without ST6GAL1 OE were treated with 100 ng/mL EGF for 15 minutes and immunoblotted for p-EGFR and t-EGFR.
    Figure Legend Snippet: A). ST6GAL1 was stably knocked-down (KD) in cells with high endogenous ST6GAL1 expression (MiaPaCa-2, S2-013, S2-LM7AA, OVCAR-3, and OVCAR-5) using lentivirus encoding an shRNA sequence targeting ST6GAL1. As controls, cells were either transduced with lentivirus containing shRNA targeting GFP (shC) or with an empty vector (EV) construct. B). Cells with undetectable endogenous ST6GAL1 (SW48 and OV4) were stably transduced with ST6GAL1-encoding cDNA to overexpress (OE) the enzyme, or with an EV construct. All cell lines represent polyclonal populations. C). Cells with or without ST6GAL1 KD were treated with 100 ng/mL EGF for 15 minutes and immunoblotted for p-EGFR (pY1068) and total EGFR (t-EGFR). D). Cells with or without ST6GAL1 OE were treated with 100 ng/mL EGF for 15 minutes and immunoblotted for p-EGFR and t-EGFR.

    Techniques Used: Stable Transfection, Expressing, shRNA, Sequencing, Transduction, Plasmid Preparation, Construct

    OVCAR-3 and OVCAR-5 cells were treated with 100 ng/mL of EGF for 10 minutes, fixed, permeabilized, stained with SNA and/or p-EGFR, and then analyzed by flow cytometry. A). Histograms depicting intracellular staining for p-EGFR before and after treatment with EGF. B). Schematic of the gating strategy. The 10% of cells with the lowest levels of α2,6 sialylation were designated as “SNA low”, and the 10% of cells with the highest levels of α2,6 sialylation were designated as “SNA high”. SNA high and SNA low cells were assessed for levels of p-EGFR. C-D). p-EGFR levels in OVCAR-3 SNA high and SNA low cells. Representative experiment in (C) and quantification in (D). E-F). p-EGFR levels in OVCAR-5 SNA high and SNA low cells. Representative experiment in (E); quantification in (F). Dotted lines indicate the highest peak of the histograms. Graphs depict the MFI +/- S.D. from three independent experiments. (not significant, ns: p > 0.05, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001) as measured by a two-way ANOVA followed by Šidák’s multiple comparison test.
    Figure Legend Snippet: OVCAR-3 and OVCAR-5 cells were treated with 100 ng/mL of EGF for 10 minutes, fixed, permeabilized, stained with SNA and/or p-EGFR, and then analyzed by flow cytometry. A). Histograms depicting intracellular staining for p-EGFR before and after treatment with EGF. B). Schematic of the gating strategy. The 10% of cells with the lowest levels of α2,6 sialylation were designated as “SNA low”, and the 10% of cells with the highest levels of α2,6 sialylation were designated as “SNA high”. SNA high and SNA low cells were assessed for levels of p-EGFR. C-D). p-EGFR levels in OVCAR-3 SNA high and SNA low cells. Representative experiment in (C) and quantification in (D). E-F). p-EGFR levels in OVCAR-5 SNA high and SNA low cells. Representative experiment in (E); quantification in (F). Dotted lines indicate the highest peak of the histograms. Graphs depict the MFI +/- S.D. from three independent experiments. (not significant, ns: p > 0.05, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001) as measured by a two-way ANOVA followed by Šidák’s multiple comparison test.

    Techniques Used: Staining, Flow Cytometry

    OV4 cells were treated with 100 ng/mL of EGF for 5, 15, or 30 minutes, or left untreated (-), and then cell lysates were immunoblotted for signaling molecules. A-B). Representative blot (A) and quantification (B) of p-EGFR and t-EGFR. C-D). Representative blot (C) and quantification (D) of p-AKT and t-AKT. E-F). Representative blot (E) and quantification (F) of p-NFκB p65 and t-NFκB p65. G-H). Representative blot (G) and quantification (H) of p-ERK1/2 and t-ERK1/2. Blots from three independent cell lysates were analyzed by densitometry and the phospho to total ratio (p/t) was calculated and normalized to β-tubulin. D.U. = densitometry units. Statistics were calculated using a two-way ANOVA followed by Šidák’s multiple comparison test. (ns: p > 0.05, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001).
    Figure Legend Snippet: OV4 cells were treated with 100 ng/mL of EGF for 5, 15, or 30 minutes, or left untreated (-), and then cell lysates were immunoblotted for signaling molecules. A-B). Representative blot (A) and quantification (B) of p-EGFR and t-EGFR. C-D). Representative blot (C) and quantification (D) of p-AKT and t-AKT. E-F). Representative blot (E) and quantification (F) of p-NFκB p65 and t-NFκB p65. G-H). Representative blot (G) and quantification (H) of p-ERK1/2 and t-ERK1/2. Blots from three independent cell lysates were analyzed by densitometry and the phospho to total ratio (p/t) was calculated and normalized to β-tubulin. D.U. = densitometry units. Statistics were calculated using a two-way ANOVA followed by Šidák’s multiple comparison test. (ns: p > 0.05, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001).

    Techniques Used:

    OVCAR-3 cells were treated with 100 ng/mL of EGF for 5, 15, or 30 minutes or left untreated (-) and then cell lysates were immunoblotted for signaling molecules. A-B). p-EGFR and t-EGFR. C-D). p-AKT and t-AKT. E-F). p-NFκB p65 and t-NFκB p65. G-H). p-ERK1/2 and t-ERK1/2. The phospho to total ratio (p/t) was calculated and normalized to β-tubulin (“Relative D.U.”). Graphs depict the mean +/- S.D. for three independent immunoblots for each signaling molecule. Statistics were calculated using a two-way ANOVA followed by Šidák’s multiple comparison test. (ns: p > 0.05, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001).
    Figure Legend Snippet: OVCAR-3 cells were treated with 100 ng/mL of EGF for 5, 15, or 30 minutes or left untreated (-) and then cell lysates were immunoblotted for signaling molecules. A-B). p-EGFR and t-EGFR. C-D). p-AKT and t-AKT. E-F). p-NFκB p65 and t-NFκB p65. G-H). p-ERK1/2 and t-ERK1/2. The phospho to total ratio (p/t) was calculated and normalized to β-tubulin (“Relative D.U.”). Graphs depict the mean +/- S.D. for three independent immunoblots for each signaling molecule. Statistics were calculated using a two-way ANOVA followed by Šidák’s multiple comparison test. (ns: p > 0.05, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001).

    Techniques Used: Western Blot

    OVCAR-5 cells were treated with 100 ng/mL of EGF for 5, 15, or 30 minutes or left untreated (-), and then cell lysates were immunoblotted for signaling molecules. A-B). p-EGFR and t-EGFR. C-D). p-AKT and t-AKT. E-F). p-NFκB p65 and t-NFκB p65. G-H). p-ERK1/2 and t-ERK1/2. The phospho to total ratio (p/t) was calculated and normalized to β-tubulin (“Relative D.U.”). Graphs depict the mean +/- S.D. for three independent immunoblots for each signaling molecule. Statistics were calculated using a two-way ANOVA followed by Šidák’s multiple comparison test. (ns: p > 0.05, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001).
    Figure Legend Snippet: OVCAR-5 cells were treated with 100 ng/mL of EGF for 5, 15, or 30 minutes or left untreated (-), and then cell lysates were immunoblotted for signaling molecules. A-B). p-EGFR and t-EGFR. C-D). p-AKT and t-AKT. E-F). p-NFκB p65 and t-NFκB p65. G-H). p-ERK1/2 and t-ERK1/2. The phospho to total ratio (p/t) was calculated and normalized to β-tubulin (“Relative D.U.”). Graphs depict the mean +/- S.D. for three independent immunoblots for each signaling molecule. Statistics were calculated using a two-way ANOVA followed by Šidák’s multiple comparison test. (ns: p > 0.05, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001).

    Techniques Used: Western Blot

    p egfr p tyr1068 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p egfr p tyr1068 antibody
    P Egfr P Tyr1068 Antibody, 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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    p egfr tyr1068  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p egfr tyr1068
    P Egfr Tyr1068, 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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    p egfr  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p egfr
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    anti p egfr  (Cell Signaling Technology Inc)


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    antibody against p egfr  (Cell Signaling Technology Inc)


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    rabbit mab anti p egfr tyr1068  (Cell Signaling Technology Inc)


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    1) Product Images from "IGFBP3 induced by the TGF-β/EGFRvIII transactivation contributes to the malignant phenotype of glioblastoma"

    Article Title: IGFBP3 induced by the TGF-β/EGFRvIII transactivation contributes to the malignant phenotype of glioblastoma

    Journal: iScience

    doi: 10.1016/j.isci.2023.106639


    Figure Legend Snippet:

    Techniques Used: Luciferase, Microarray, Recombinant, Lysis, Transfection, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, Reporter Gene Assay, BIA-KA, Cell Cycle Assay, Sequencing, shRNA, Real-time Polymerase Chain Reaction, Plasmid Preparation, Software

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    Journal: iScience

    Article Title: IGFBP3 induced by the TGF-β/EGFRvIII transactivation contributes to the malignant phenotype of glioblastoma

    doi: 10.1016/j.isci.2023.106639

    Figure Lengend Snippet:

    Article Snippet: Rabbit mAb anti-p-EGFR (Tyr1068) (D7A5) , Cell Signaling Technology , Cat#3777; RRID: AB_2096270.

    Techniques: Luciferase, Microarray, Recombinant, Lysis, Transfection, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, Reporter Gene Assay, BIA-KA, Cell Cycle Assay, Sequencing, shRNA, Real-time Polymerase Chain Reaction, Plasmid Preparation, Software