egfr  (Abcam)

 
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  • 96
    Name:
    Anti EGFR antibody EP38Y
    Description:

    Catalog Number:
    AB52894
    Price:
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    Structured Review

    Abcam egfr
    Calcium chelator inhibits <t>calpain</t> activity and ITGβ4 cleavage in LMP2A-expressing NPC cells. (A) Calpain activity measured in a fluorometric assay in LMP2A-positive CNE1/TW03 cell lines before and after treatment with BAPTA-AM (10 μm/l) for 24 h. Data are mean±s.d. ( n =3). (B) Top panel: immunoblots showing the expression of ITGβ4 (205 kDa), cleaved ITGβ4 isoforms (165/125 kDa) and tyrosine-phosphorylated <t>EGFR</t> in LMP2A-expressing and control cells. GAPDH was used as loading control. Bottom panel: semi-quantitative analysis of three repeated immunoblot experiments as in the top panel. Relative expression of the cleaved ITGβ4 isoforms (left) and pEGFR (right) under the same experimental conditions. ** P

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    Images

    1) Product Images from "Epstein-Barr virus-encoded LMP2A stimulates migration of nasopharyngeal carcinoma cells via the EGFR/Ca2+/calpain/ITGβ4 axis"

    Article Title: Epstein-Barr virus-encoded LMP2A stimulates migration of nasopharyngeal carcinoma cells via the EGFR/Ca2+/calpain/ITGβ4 axis

    Journal: Biology Open

    doi: 10.1242/bio.024646

    Calcium chelator inhibits calpain activity and ITGβ4 cleavage in LMP2A-expressing NPC cells. (A) Calpain activity measured in a fluorometric assay in LMP2A-positive CNE1/TW03 cell lines before and after treatment with BAPTA-AM (10 μm/l) for 24 h. Data are mean±s.d. ( n =3). (B) Top panel: immunoblots showing the expression of ITGβ4 (205 kDa), cleaved ITGβ4 isoforms (165/125 kDa) and tyrosine-phosphorylated EGFR in LMP2A-expressing and control cells. GAPDH was used as loading control. Bottom panel: semi-quantitative analysis of three repeated immunoblot experiments as in the top panel. Relative expression of the cleaved ITGβ4 isoforms (left) and pEGFR (right) under the same experimental conditions. ** P
    Figure Legend Snippet: Calcium chelator inhibits calpain activity and ITGβ4 cleavage in LMP2A-expressing NPC cells. (A) Calpain activity measured in a fluorometric assay in LMP2A-positive CNE1/TW03 cell lines before and after treatment with BAPTA-AM (10 μm/l) for 24 h. Data are mean±s.d. ( n =3). (B) Top panel: immunoblots showing the expression of ITGβ4 (205 kDa), cleaved ITGβ4 isoforms (165/125 kDa) and tyrosine-phosphorylated EGFR in LMP2A-expressing and control cells. GAPDH was used as loading control. Bottom panel: semi-quantitative analysis of three repeated immunoblot experiments as in the top panel. Relative expression of the cleaved ITGβ4 isoforms (left) and pEGFR (right) under the same experimental conditions. ** P

    Techniques Used: Activity Assay, Expressing, Western Blot

    2) Product Images from "Analyses of Intravesicular Exosomal Proteins Using a Nano-Plasmonic System"

    Article Title: Analyses of Intravesicular Exosomal Proteins Using a Nano-Plasmonic System

    Journal: ACS photonics

    doi: 10.1021/acsphotonics.7b00992

    Drug response monitoring by EV protein analysis (a) OV90 cells were treated with HSP90 inhibitor (17-AAG, 10 μ M) for 48 h in conditioned media. EVs secreted from OV90 were collected and the lysates were analyzed by iNPS (10 8 EVs per marker). Protein level fold changes after drug treatment were calculated from iNPS spectral shifts which were proportional to the amount of target proteins captured on the sensing surface. We observed the increase of HPS90 and HSP70, and the decrease of AKT1, which reflects the known working mechanism of the inhibitor. (b) 17-AAG mediated protein expression change of OV90 cell and EVs were monitored vi a western blotting. EV protein profiles matched with iNPS results. (c) OV429 cells were treated with 20 μM EGFR inhibitor (gefitinib) for 48 h in conditioned media, and EVs were analyzed by iNPS. Significant increases in EGFR, EpCAM and CD63 were observed with EVs from drug-treated cells. The error bars represent the standard deviation of signals. (d) Gefitinib mediated protein expression change of OV429 cell and EVs were monitored vi a western blotting. Note the expression differences between cells and EVs upon treatment.
    Figure Legend Snippet: Drug response monitoring by EV protein analysis (a) OV90 cells were treated with HSP90 inhibitor (17-AAG, 10 μ M) for 48 h in conditioned media. EVs secreted from OV90 were collected and the lysates were analyzed by iNPS (10 8 EVs per marker). Protein level fold changes after drug treatment were calculated from iNPS spectral shifts which were proportional to the amount of target proteins captured on the sensing surface. We observed the increase of HPS90 and HSP70, and the decrease of AKT1, which reflects the known working mechanism of the inhibitor. (b) 17-AAG mediated protein expression change of OV90 cell and EVs were monitored vi a western blotting. EV protein profiles matched with iNPS results. (c) OV429 cells were treated with 20 μM EGFR inhibitor (gefitinib) for 48 h in conditioned media, and EVs were analyzed by iNPS. Significant increases in EGFR, EpCAM and CD63 were observed with EVs from drug-treated cells. The error bars represent the standard deviation of signals. (d) Gefitinib mediated protein expression change of OV429 cell and EVs were monitored vi a western blotting. Note the expression differences between cells and EVs upon treatment.

    Techniques Used: Marker, Expressing, Western Blot, Standard Deviation

    3) Product Images from "miR-206 inhibits liver cancer stem cell expansion by regulating EGFR expression"

    Article Title: miR-206 inhibits liver cancer stem cell expansion by regulating EGFR expression

    Journal: Cell Cycle

    doi: 10.1080/15384101.2020.1739808

    EGFR was a direct target of miR-206 in liver CSCs. (a). The protein expression of EGFR was checked in Huh7 miR-206 or HCCLM3 miR-206 and their control cells by western blot. GAPDH acted as a loading control. (b). The mRNA expression of EGFR was checked in Huh7 miR-206 or HCCLM3 miR-206 and their control cells by RT-PCR. (n = 3). (c). Luciferase reporter assays performed in Huh7 miR-206 or HCCLM3 miR-206 and their control cells transfected with EGFR 3ʹ-UTR constructs. (n = 3). (d). Luciferase reporter assays performed in Huh7 miR-206 or HCCLM3 miR-206 and their control cells transfected with wild-type or mutant EGFR 3ʹ-UTR constructs. (n = 3). (e). Huh7 miR-206 and its control cells were treated with Gefitinib (10 μM) or not and then checked by flow-cytometric assay. (n = 3). (f). Huh7 miR-206 or HCCLM3 miR-206 and their control cells were treated with Gefitinib (10 μM) or not and then subjected spheres formation assay. (n = 4). (g). Spearman correlation analysis of the relationship between EGFR protein and miR-206 expression in 40 HCC tissues.(Data are represented as mean±s.d.; *P
    Figure Legend Snippet: EGFR was a direct target of miR-206 in liver CSCs. (a). The protein expression of EGFR was checked in Huh7 miR-206 or HCCLM3 miR-206 and their control cells by western blot. GAPDH acted as a loading control. (b). The mRNA expression of EGFR was checked in Huh7 miR-206 or HCCLM3 miR-206 and their control cells by RT-PCR. (n = 3). (c). Luciferase reporter assays performed in Huh7 miR-206 or HCCLM3 miR-206 and their control cells transfected with EGFR 3ʹ-UTR constructs. (n = 3). (d). Luciferase reporter assays performed in Huh7 miR-206 or HCCLM3 miR-206 and their control cells transfected with wild-type or mutant EGFR 3ʹ-UTR constructs. (n = 3). (e). Huh7 miR-206 and its control cells were treated with Gefitinib (10 μM) or not and then checked by flow-cytometric assay. (n = 3). (f). Huh7 miR-206 or HCCLM3 miR-206 and their control cells were treated with Gefitinib (10 μM) or not and then subjected spheres formation assay. (n = 4). (g). Spearman correlation analysis of the relationship between EGFR protein and miR-206 expression in 40 HCC tissues.(Data are represented as mean±s.d.; *P

    Techniques Used: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Luciferase, Transfection, Construct, Mutagenesis, Flow Cytometry, Tube Formation Assay

    4) Product Images from "Differential Expression of RBM5, EGFR and KRAS mRNA and protein in non-small cell lung cancer tissues"

    Article Title: Differential Expression of RBM5, EGFR and KRAS mRNA and protein in non-small cell lung cancer tissues

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/1756-9966-31-36

    Expression of RBM5, EGFR and KRAS protein in NSCLC . A, Western blot of RBM5, EGFR and KRAS protein expression in representative tissue samples from NSCLC and non-tumor specimens. Total cellular protein was extracted, subjected to Western blot analysis and quantified using Quantity One software. B, Quantitative data from A. *p
    Figure Legend Snippet: Expression of RBM5, EGFR and KRAS protein in NSCLC . A, Western blot of RBM5, EGFR and KRAS protein expression in representative tissue samples from NSCLC and non-tumor specimens. Total cellular protein was extracted, subjected to Western blot analysis and quantified using Quantity One software. B, Quantitative data from A. *p

    Techniques Used: Expressing, Western Blot, Software

    Expression of RBM5, EGFR and KRAS mRNA in NSCLC . A, Agarose gel of semi-quantitative RT-PCR data of RBM5, EGFR, and KRAS mRNA expression in representative NSCLC and non-tumor specimens. Total RNA was isolated and subjected to semi-quantitative RT-PCR and quantified using Quantity One software. B , Quantitative data from A. *p
    Figure Legend Snippet: Expression of RBM5, EGFR and KRAS mRNA in NSCLC . A, Agarose gel of semi-quantitative RT-PCR data of RBM5, EGFR, and KRAS mRNA expression in representative NSCLC and non-tumor specimens. Total RNA was isolated and subjected to semi-quantitative RT-PCR and quantified using Quantity One software. B , Quantitative data from A. *p

    Techniques Used: Expressing, Agarose Gel Electrophoresis, Quantitative RT-PCR, Isolation, Software

    5) Product Images from "A murine model of dry eye induced by topical administration of erlotinib eye drops"

    Article Title: A murine model of dry eye induced by topical administration of erlotinib eye drops

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2017.3353

    Effect of erlotinib on TNF-α, EGFR and p-EGFR protein expression levels. (A) Following treatment for 3 weeks with either erlotinib or PBS, the protein expression levels of TNF-α were determined by western blot analysis, and the (B) density of the bands was statistically analyzed. (C) Following treatment for 3 weeks with either erlotinib or PBS, the protein expression levels of EGFR and p-EGFR were determined by western blot analysis, and the (D) density of the bands was used to statistically analyze the p-EGFR/EGFR ratio. Data are presented as mean + standard deviation. * P
    Figure Legend Snippet: Effect of erlotinib on TNF-α, EGFR and p-EGFR protein expression levels. (A) Following treatment for 3 weeks with either erlotinib or PBS, the protein expression levels of TNF-α were determined by western blot analysis, and the (B) density of the bands was statistically analyzed. (C) Following treatment for 3 weeks with either erlotinib or PBS, the protein expression levels of EGFR and p-EGFR were determined by western blot analysis, and the (D) density of the bands was used to statistically analyze the p-EGFR/EGFR ratio. Data are presented as mean + standard deviation. * P

    Techniques Used: Expressing, Western Blot, Standard Deviation

    6) Product Images from "EGFR expression is associated with cytoplasmic staining of CXCR4 and predicts poor prognosis in triple-negative breast carcinomas"

    Article Title: EGFR expression is associated with cytoplasmic staining of CXCR4 and predicts poor prognosis in triple-negative breast carcinomas

    Journal: Oncology Letters

    doi: 10.3892/ol.2016.5489

    Immunohistochemical staining of (A-D) EGFR and (E-H) CXCR4 expression in breast cancer. (A) Negative expression (−) of EGFR; (B) weak expression (+) of EGFR; (C) moderate expression (++) of EGFR; (D) intense expression (+++) of EGFR; (E) negative expression (−) of CXCR4; (F) weak expression (+) of CXCR4; (G) moderate expression (++) of CXCR4; (H) intense expression (+++) of CXCR4. Magnification, 400x; scale bar, 100 µm. EGFR, epidermal growth factor receptor; CXCR4, C-X-C motif chemokine receptor type 4.
    Figure Legend Snippet: Immunohistochemical staining of (A-D) EGFR and (E-H) CXCR4 expression in breast cancer. (A) Negative expression (−) of EGFR; (B) weak expression (+) of EGFR; (C) moderate expression (++) of EGFR; (D) intense expression (+++) of EGFR; (E) negative expression (−) of CXCR4; (F) weak expression (+) of CXCR4; (G) moderate expression (++) of CXCR4; (H) intense expression (+++) of CXCR4. Magnification, 400x; scale bar, 100 µm. EGFR, epidermal growth factor receptor; CXCR4, C-X-C motif chemokine receptor type 4.

    Techniques Used: Immunohistochemistry, Staining, Expressing

    Downregulation of CXCR4 and/or EGFR by shRNA in MDA-MB-231 cells inhibits xenograft tumor growth in mouse models. (A) A total of 2×10 6 stably transduced MDA-MB-231 cells were implanted into the right legs of nude mice. The mice (n=3 per group) were observed for 4 weeks. (B) Tumors were measured every week with external calipers and tumor volume was calculated according to the following formula: Volume = 0.5 × a 2 × b, where ‘a’ is the smallest superficial diameter and ‘b’ is the largest superficial diameter. (C) After 4 weeks, the tumors were excised and weighed. *P
    Figure Legend Snippet: Downregulation of CXCR4 and/or EGFR by shRNA in MDA-MB-231 cells inhibits xenograft tumor growth in mouse models. (A) A total of 2×10 6 stably transduced MDA-MB-231 cells were implanted into the right legs of nude mice. The mice (n=3 per group) were observed for 4 weeks. (B) Tumors were measured every week with external calipers and tumor volume was calculated according to the following formula: Volume = 0.5 × a 2 × b, where ‘a’ is the smallest superficial diameter and ‘b’ is the largest superficial diameter. (C) After 4 weeks, the tumors were excised and weighed. *P

    Techniques Used: shRNA, Multiple Displacement Amplification, Stable Transfection, Mouse Assay

    EGFR and CXCR4 expression levels are reciprocally affected in MDA-MB-231 cells, and alter the biological behavior of cells. (A) Western blot analysis results: (a) Transfection with shEGFR #1 effectively blocked the expression of EGFR protein; (b) shCXCR4 #2 blocked the expression of CXCR4 protein almost completely; (c) when EGFR was suppressed, CXCR4 was reduced. Each experiment was repeated ≥3 times. (B) The MTT assay results revealed that MDA-MB-231 cells with suppressed CXCR4 or EGFR exhibited a significant reduction in proliferation, compared with those transfected with the empty vector control, and the greatest inhibition was observed in cells in which EGFR and CXCR4 were both suppressed. (C) Transwell migration and (D) Transwell invasion assays were performed in MDA-MB-231 cells with EGFR and/or CXCR4 silenced by shRNA, and the results were quantified by comparison with the empty vector-transfected MDA-MB-231 cells. *P
    Figure Legend Snippet: EGFR and CXCR4 expression levels are reciprocally affected in MDA-MB-231 cells, and alter the biological behavior of cells. (A) Western blot analysis results: (a) Transfection with shEGFR #1 effectively blocked the expression of EGFR protein; (b) shCXCR4 #2 blocked the expression of CXCR4 protein almost completely; (c) when EGFR was suppressed, CXCR4 was reduced. Each experiment was repeated ≥3 times. (B) The MTT assay results revealed that MDA-MB-231 cells with suppressed CXCR4 or EGFR exhibited a significant reduction in proliferation, compared with those transfected with the empty vector control, and the greatest inhibition was observed in cells in which EGFR and CXCR4 were both suppressed. (C) Transwell migration and (D) Transwell invasion assays were performed in MDA-MB-231 cells with EGFR and/or CXCR4 silenced by shRNA, and the results were quantified by comparison with the empty vector-transfected MDA-MB-231 cells. *P

    Techniques Used: Expressing, Multiple Displacement Amplification, Western Blot, Transfection, MTT Assay, Plasmid Preparation, Inhibition, Migration, shRNA

    7) Product Images from "EGFR expression is associated with cytoplasmic staining of CXCR4 and predicts poor prognosis in triple-negative breast carcinomas"

    Article Title: EGFR expression is associated with cytoplasmic staining of CXCR4 and predicts poor prognosis in triple-negative breast carcinomas

    Journal: Oncology Letters

    doi: 10.3892/ol.2016.5489

    Immunohistochemical staining of (A-D) EGFR and (E-H) CXCR4 expression in breast cancer. (A) Negative expression (−) of EGFR; (B) weak expression (+) of EGFR; (C) moderate expression (++) of EGFR; (D) intense expression (+++) of EGFR; (E) negative expression (−) of CXCR4; (F) weak expression (+) of CXCR4; (G) moderate expression (++) of CXCR4; (H) intense expression (+++) of CXCR4. Magnification, 400x; scale bar, 100 µm. EGFR, epidermal growth factor receptor; CXCR4, C-X-C motif chemokine receptor type 4.
    Figure Legend Snippet: Immunohistochemical staining of (A-D) EGFR and (E-H) CXCR4 expression in breast cancer. (A) Negative expression (−) of EGFR; (B) weak expression (+) of EGFR; (C) moderate expression (++) of EGFR; (D) intense expression (+++) of EGFR; (E) negative expression (−) of CXCR4; (F) weak expression (+) of CXCR4; (G) moderate expression (++) of CXCR4; (H) intense expression (+++) of CXCR4. Magnification, 400x; scale bar, 100 µm. EGFR, epidermal growth factor receptor; CXCR4, C-X-C motif chemokine receptor type 4.

    Techniques Used: Immunohistochemistry, Staining, Expressing

    Downregulation of CXCR4 and/or EGFR by shRNA in MDA-MB-231 cells inhibits xenograft tumor growth in mouse models. (A) A total of 2×10 6 stably transduced MDA-MB-231 cells were implanted into the right legs of nude mice. The mice (n=3 per group) were observed for 4 weeks. (B) Tumors were measured every week with external calipers and tumor volume was calculated according to the following formula: Volume = 0.5 × a 2 × b, where ‘a’ is the smallest superficial diameter and ‘b’ is the largest superficial diameter. (C) After 4 weeks, the tumors were excised and weighed. *P
    Figure Legend Snippet: Downregulation of CXCR4 and/or EGFR by shRNA in MDA-MB-231 cells inhibits xenograft tumor growth in mouse models. (A) A total of 2×10 6 stably transduced MDA-MB-231 cells were implanted into the right legs of nude mice. The mice (n=3 per group) were observed for 4 weeks. (B) Tumors were measured every week with external calipers and tumor volume was calculated according to the following formula: Volume = 0.5 × a 2 × b, where ‘a’ is the smallest superficial diameter and ‘b’ is the largest superficial diameter. (C) After 4 weeks, the tumors were excised and weighed. *P

    Techniques Used: shRNA, Multiple Displacement Amplification, Stable Transfection, Mouse Assay

    EGFR and CXCR4 expression levels are reciprocally affected in MDA-MB-231 cells, and alter the biological behavior of cells. (A) Western blot analysis results: (a) Transfection with shEGFR #1 effectively blocked the expression of EGFR protein; (b) shCXCR4 #2 blocked the expression of CXCR4 protein almost completely; (c) when EGFR was suppressed, CXCR4 was reduced. Each experiment was repeated ≥3 times. (B) The MTT assay results revealed that MDA-MB-231 cells with suppressed CXCR4 or EGFR exhibited a significant reduction in proliferation, compared with those transfected with the empty vector control, and the greatest inhibition was observed in cells in which EGFR and CXCR4 were both suppressed. (C) Transwell migration and (D) Transwell invasion assays were performed in MDA-MB-231 cells with EGFR and/or CXCR4 silenced by shRNA, and the results were quantified by comparison with the empty vector-transfected MDA-MB-231 cells. *P
    Figure Legend Snippet: EGFR and CXCR4 expression levels are reciprocally affected in MDA-MB-231 cells, and alter the biological behavior of cells. (A) Western blot analysis results: (a) Transfection with shEGFR #1 effectively blocked the expression of EGFR protein; (b) shCXCR4 #2 blocked the expression of CXCR4 protein almost completely; (c) when EGFR was suppressed, CXCR4 was reduced. Each experiment was repeated ≥3 times. (B) The MTT assay results revealed that MDA-MB-231 cells with suppressed CXCR4 or EGFR exhibited a significant reduction in proliferation, compared with those transfected with the empty vector control, and the greatest inhibition was observed in cells in which EGFR and CXCR4 were both suppressed. (C) Transwell migration and (D) Transwell invasion assays were performed in MDA-MB-231 cells with EGFR and/or CXCR4 silenced by shRNA, and the results were quantified by comparison with the empty vector-transfected MDA-MB-231 cells. *P

    Techniques Used: Expressing, Multiple Displacement Amplification, Western Blot, Transfection, MTT Assay, Plasmid Preparation, Inhibition, Migration, shRNA

    8) Product Images from "Ectodysplasin A protein promotes corneal epithelial cell proliferation"

    Article Title: Ectodysplasin A protein promotes corneal epithelial cell proliferation

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M117.803809

    Eda activated EGF-EGFR signaling pathway in HCE cells. A, HCE cells were cultured with different concentrations of Eda (0, 5, 10, and 20 ng/ml). CCK8 assay results showed 5 and 10 ng/ml of Eda-enhanced cell proliferation, and reached a plateau at 10 ng/ml ( n = 4, **, p
    Figure Legend Snippet: Eda activated EGF-EGFR signaling pathway in HCE cells. A, HCE cells were cultured with different concentrations of Eda (0, 5, 10, and 20 ng/ml). CCK8 assay results showed 5 and 10 ng/ml of Eda-enhanced cell proliferation, and reached a plateau at 10 ng/ml ( n = 4, **, p

    Techniques Used: Cell Culture, CCK-8 Assay

    9) Product Images from "Positive feedback of the amphiregulin-EGFR-ERK pathway mediates PM2.5 from wood smoke-induced MUC5AC expression in epithelial cells"

    Article Title: Positive feedback of the amphiregulin-EGFR-ERK pathway mediates PM2.5 from wood smoke-induced MUC5AC expression in epithelial cells

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-11541-1

    Ligand-dependent EGFR activation mediates MUC5AC induction by WSPM2.5. ( A ) NCI-H292 cells were stimulated with WSPM2.5 (8 μg/ml) for the indicated times. Cell lysates were prepared for Western blot analysis. Antibodies specific for EGFR, phosphorylated EGFR (p-EGFR) and GAPDH were used. ( B ) Density quantification of EGFR phosphorylation in ( A ). ( C ) NCI-H292 cells were pretreated with or without AG1478 (5, 10, or 20 μM) for 1 h and were then treated with WSPM2.5 for 24 h to measure MUC5AC mRNA expression. ( D ) NCI-H292 cells were pretreated with or without an EGFR-neutralizing antibody or a mouse control IgG antibody (0.5 μg/ml) for 1 h and were then stimulated with WSPM2.5 (8 μg/ml) for 60 min. Cell lysates were collected to examine EGFR phosphorylation. ( E ) Density quantification of EGFR phosphorylation in ( D ). ( F ) NCI-H292 cells were pretreated with or without different concentrations of EGFR-neutralizing antibody (0.5, 1, or 2 μg/ml) for 1 h before treatment with WSPM2.5 (8 μg/ml) for 24 h to analyze the MUC5AC mRNA expression levels. ( A and D ) one representative gel is shown. Densitometry analysis of p-EGFR was performed after normalization to GAPDH. The data are expressed as the means ± SD. n = 5 ( A,B and C ) or n = 4 ( D , E and F ). * P
    Figure Legend Snippet: Ligand-dependent EGFR activation mediates MUC5AC induction by WSPM2.5. ( A ) NCI-H292 cells were stimulated with WSPM2.5 (8 μg/ml) for the indicated times. Cell lysates were prepared for Western blot analysis. Antibodies specific for EGFR, phosphorylated EGFR (p-EGFR) and GAPDH were used. ( B ) Density quantification of EGFR phosphorylation in ( A ). ( C ) NCI-H292 cells were pretreated with or without AG1478 (5, 10, or 20 μM) for 1 h and were then treated with WSPM2.5 for 24 h to measure MUC5AC mRNA expression. ( D ) NCI-H292 cells were pretreated with or without an EGFR-neutralizing antibody or a mouse control IgG antibody (0.5 μg/ml) for 1 h and were then stimulated with WSPM2.5 (8 μg/ml) for 60 min. Cell lysates were collected to examine EGFR phosphorylation. ( E ) Density quantification of EGFR phosphorylation in ( D ). ( F ) NCI-H292 cells were pretreated with or without different concentrations of EGFR-neutralizing antibody (0.5, 1, or 2 μg/ml) for 1 h before treatment with WSPM2.5 (8 μg/ml) for 24 h to analyze the MUC5AC mRNA expression levels. ( A and D ) one representative gel is shown. Densitometry analysis of p-EGFR was performed after normalization to GAPDH. The data are expressed as the means ± SD. n = 5 ( A,B and C ) or n = 4 ( D , E and F ). * P

    Techniques Used: Activation Assay, Western Blot, Expressing

    AR is the primary ligand responsible for the activation of EGFR-ERK signaling and MUC5AC expression. Cells were pretreated with neutralizing antibodies against AR, TGF-α or HB-EGF for 1 h and were then stimulated with WSPM2.5 for 60 or 5 min to assay for phosphorylation of EGFR and ERK, respectively ( A ), or for 24 h to measure MUC5AC mRNA expression ( D ). ( B ) Density quantification of EGFR phosphorylation, and ( C ) density quantification of ERK phosphorylation in ( A ). ( E ) NCI-H292 cells were exposed to recombinant AR (300–1,800 pg/ml) for 24 h to assess MUC5AC mRNA expression. ( A ) one representative gel from three independent experiments is shown. Densitometry analysis of p-EGFR and p-ERK after normalization to GAPDH or total ERK, respectively. The data are expressed as the means ± SD; n = 3. * P
    Figure Legend Snippet: AR is the primary ligand responsible for the activation of EGFR-ERK signaling and MUC5AC expression. Cells were pretreated with neutralizing antibodies against AR, TGF-α or HB-EGF for 1 h and were then stimulated with WSPM2.5 for 60 or 5 min to assay for phosphorylation of EGFR and ERK, respectively ( A ), or for 24 h to measure MUC5AC mRNA expression ( D ). ( B ) Density quantification of EGFR phosphorylation, and ( C ) density quantification of ERK phosphorylation in ( A ). ( E ) NCI-H292 cells were exposed to recombinant AR (300–1,800 pg/ml) for 24 h to assess MUC5AC mRNA expression. ( A ) one representative gel from three independent experiments is shown. Densitometry analysis of p-EGFR and p-ERK after normalization to GAPDH or total ERK, respectively. The data are expressed as the means ± SD; n = 3. * P

    Techniques Used: Activation Assay, Expressing, Recombinant

    WSPM2.5-induced MUC5AC expression is mediated through the EGFR-dependent ERK pathway. ( A ) NCI-H292 cells were stimulated with WSPM2.5 for the indicated durations. Cell lysates were collected and assayed for ERK phosphorylation. The membranes were stripped and re-probed with an anti-ERK antibody. ( B ) Density quantification of ERK phosphorylation in ( A ). ( C ) NCI-H292 cells were pretreated with or without PD98059 (5, 10, or 20 µM) for 1 h and were then treated with WSPM2.5 for 24 h. MUC5AC mRNA expression was determined by real-time PCR. ( D ) NCI-H292 cells were pretreated with or without AG1478 (5, 10, or 20 μM) for 1 h and were then stimulated with WSPM2.5 for 5 min to assay for ERK phosphorylation. ( E ) Density quantification of ERK phosphorylation in ( D ). ( A and D ) one representative gel from three independent experiments is shown. The data are expressed as the means ± SD (n = 3). * P
    Figure Legend Snippet: WSPM2.5-induced MUC5AC expression is mediated through the EGFR-dependent ERK pathway. ( A ) NCI-H292 cells were stimulated with WSPM2.5 for the indicated durations. Cell lysates were collected and assayed for ERK phosphorylation. The membranes were stripped and re-probed with an anti-ERK antibody. ( B ) Density quantification of ERK phosphorylation in ( A ). ( C ) NCI-H292 cells were pretreated with or without PD98059 (5, 10, or 20 µM) for 1 h and were then treated with WSPM2.5 for 24 h. MUC5AC mRNA expression was determined by real-time PCR. ( D ) NCI-H292 cells were pretreated with or without AG1478 (5, 10, or 20 μM) for 1 h and were then stimulated with WSPM2.5 for 5 min to assay for ERK phosphorylation. ( E ) Density quantification of ERK phosphorylation in ( D ). ( A and D ) one representative gel from three independent experiments is shown. The data are expressed as the means ± SD (n = 3). * P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    10) Product Images from "Evidence That ADAM17 Mediates the Protective Action of CGRP against Angiotensin II-Induced Inflammation in Vascular Smooth Muscle Cells"

    Article Title: Evidence That ADAM17 Mediates the Protective Action of CGRP against Angiotensin II-Induced Inflammation in Vascular Smooth Muscle Cells

    Journal: Mediators of Inflammation

    doi: 10.1155/2018/2109352

    EGFR activation is required in Ang II-induced inflammation in vascular smooth muscle cells. After being pretreated with selective EGFR inhibitor AG1478 (5 μ mol/l) for 30 minutes, vascular smooth muscle cells were then stimulated with 100 nmol/l Ang II for 24 hours. (a) EGFR activity, n = 4 independent experiments. (b) ERK1/2 activity, n = 3 independent experiments. (c–e) At the end of the experiment, the medium was collected to detect the concentration of IL-1 β , IL-6, and TNF- α by ELISA, n = 3–5 independent experiments. (c) IL-1 β protein level. (d) IL-6 protein level. (e) TNF- α protein level. (f) The mRNA levels of IL-1 β , IL-6, and TNF- α , n = 3 − 4 independent experiments. Ang II represents angiotensin II. ∗ P
    Figure Legend Snippet: EGFR activation is required in Ang II-induced inflammation in vascular smooth muscle cells. After being pretreated with selective EGFR inhibitor AG1478 (5 μ mol/l) for 30 minutes, vascular smooth muscle cells were then stimulated with 100 nmol/l Ang II for 24 hours. (a) EGFR activity, n = 4 independent experiments. (b) ERK1/2 activity, n = 3 independent experiments. (c–e) At the end of the experiment, the medium was collected to detect the concentration of IL-1 β , IL-6, and TNF- α by ELISA, n = 3–5 independent experiments. (c) IL-1 β protein level. (d) IL-6 protein level. (e) TNF- α protein level. (f) The mRNA levels of IL-1 β , IL-6, and TNF- α , n = 3 − 4 independent experiments. Ang II represents angiotensin II. ∗ P

    Techniques Used: Activation Assay, Activity Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay

    CGRP reduced ADAM17 expression and activation of EGFR and ERK1/2 in vascular smooth muscle cells treated with Ang II. After being pretreated with 10 nmol/l CGRP or/and 10 nmol/l CGRP 8–37 for 30 minutes, vascular smooth muscle cells were treated with 100 nmol/l Ang II for 24 hours. (a) ADAM17 protein level, n = 4 independent experiments. (b) ADAM17 mRNA level, n = 3 independent experiments. (c) EGFR activity, n = 4 independent experiments. (d) ERK1/2 activity, n = 3 independent experiments. Ang II represents angiotensin II. ∗ P
    Figure Legend Snippet: CGRP reduced ADAM17 expression and activation of EGFR and ERK1/2 in vascular smooth muscle cells treated with Ang II. After being pretreated with 10 nmol/l CGRP or/and 10 nmol/l CGRP 8–37 for 30 minutes, vascular smooth muscle cells were treated with 100 nmol/l Ang II for 24 hours. (a) ADAM17 protein level, n = 4 independent experiments. (b) ADAM17 mRNA level, n = 3 independent experiments. (c) EGFR activity, n = 4 independent experiments. (d) ERK1/2 activity, n = 3 independent experiments. Ang II represents angiotensin II. ∗ P

    Techniques Used: Expressing, Activation Assay, Activity Assay

    ADAM17 siRNA decreased Ang II-induced activation of EGFR and ERK1/2 in vascular smooth muscle cells. After being transfected with 100 nmol/l ADAM17 siRNA for 30 hours, vascular smooth muscle cells were then stimulated with 100 nmol/l Ang II for 24 hours. (a) EGFR activity, n = 4 independent experiments; (b) ERK1/2 activity, n = 3 independent experiments; (c) proposed model of ADAM17 in regulating the inhibitory effect of CGRP on Ang II-induced inflammation through the EGFR-ERK1/2 pathway. Ang II: angiotensin II; NC: negative control; +: positive effect; −: negative effect; VSMCs: vascular smooth muscle cells. ∗ P
    Figure Legend Snippet: ADAM17 siRNA decreased Ang II-induced activation of EGFR and ERK1/2 in vascular smooth muscle cells. After being transfected with 100 nmol/l ADAM17 siRNA for 30 hours, vascular smooth muscle cells were then stimulated with 100 nmol/l Ang II for 24 hours. (a) EGFR activity, n = 4 independent experiments; (b) ERK1/2 activity, n = 3 independent experiments; (c) proposed model of ADAM17 in regulating the inhibitory effect of CGRP on Ang II-induced inflammation through the EGFR-ERK1/2 pathway. Ang II: angiotensin II; NC: negative control; +: positive effect; −: negative effect; VSMCs: vascular smooth muscle cells. ∗ P

    Techniques Used: Activation Assay, Transfection, Activity Assay, Negative Control

    11) Product Images from "Nuclear Epidermal Growth Factor Receptor is a Functional Molecular Target in Triple-Negative Breast Cancer"

    Article Title: Nuclear Epidermal Growth Factor Receptor is a Functional Molecular Target in Triple-Negative Breast Cancer

    Journal: Molecular cancer therapeutics

    doi: 10.1158/1535-7163.MCT-13-1021

    Src Family Kinases mediate nEGFR translocation in TNBC (A) Constitutively active Src (caSrc) enhances nEGFR translocation in TNBC cell lines . Cells were transfected with caSrc or an empty vector control for 48 hr prior to stimulation with EGF (5 nM, 45 min) to induce nEGFR translocation. Non-nuclear and nuclear proteins were harvested. nEGFR expression was quantitated using ImageJ software (B) A negative regulator of Src, src-like adaptor protein (SLAP), blocks nEGFR translocation in TNBC cell lines. Cells were transfected with SLAP-FLAG or an empty vector control for 48 hr prior to harvesting non-nuclear and nuclear proteins. nEGFR expression was analyzed. (C) SLAP can interact with EGFR and decrease EGFR activation at Tyrosine 1101. Cells were transfected with SLAP-FLAG or an empty vector control for 48 hr prior to harvesting whole cell lysate. 250 ug of cell lysate was immunoprecipitated with an anti-SLAP antibody. The same lysate was subjected to immunoblot analysis for activation of EGFR at Tyrosine 1101. pEGFR-Y1101 activity was quantitated using ImageJ software. Inset 1: EGFR mutated at Tyrosine 1101 is deficient in nuclear localization. Vector, EGFR-WT, and EGFR-Y1101F were transfected into CHOK1 cells for 48 hr prior to stimulation with EGF (5 nM, 45 min). Non-nuclear and nuclear proteins were harvested, and nEGFR expression was analyzed.
    Figure Legend Snippet: Src Family Kinases mediate nEGFR translocation in TNBC (A) Constitutively active Src (caSrc) enhances nEGFR translocation in TNBC cell lines . Cells were transfected with caSrc or an empty vector control for 48 hr prior to stimulation with EGF (5 nM, 45 min) to induce nEGFR translocation. Non-nuclear and nuclear proteins were harvested. nEGFR expression was quantitated using ImageJ software (B) A negative regulator of Src, src-like adaptor protein (SLAP), blocks nEGFR translocation in TNBC cell lines. Cells were transfected with SLAP-FLAG or an empty vector control for 48 hr prior to harvesting non-nuclear and nuclear proteins. nEGFR expression was analyzed. (C) SLAP can interact with EGFR and decrease EGFR activation at Tyrosine 1101. Cells were transfected with SLAP-FLAG or an empty vector control for 48 hr prior to harvesting whole cell lysate. 250 ug of cell lysate was immunoprecipitated with an anti-SLAP antibody. The same lysate was subjected to immunoblot analysis for activation of EGFR at Tyrosine 1101. pEGFR-Y1101 activity was quantitated using ImageJ software. Inset 1: EGFR mutated at Tyrosine 1101 is deficient in nuclear localization. Vector, EGFR-WT, and EGFR-Y1101F were transfected into CHOK1 cells for 48 hr prior to stimulation with EGF (5 nM, 45 min). Non-nuclear and nuclear proteins were harvested, and nEGFR expression was analyzed.

    Techniques Used: Translocation Assay, Transfection, Plasmid Preparation, Expressing, Software, Activation Assay, Immunoprecipitation, Activity Assay

    12) Product Images from "Genomic and Molecular Characterization of Malignant Peripheral Nerve Sheath Tumor Identifies the IGF1R Pathway as a Primary Target for Treatment"

    Article Title: Genomic and Molecular Characterization of Malignant Peripheral Nerve Sheath Tumor Identifies the IGF1R Pathway as a Primary Target for Treatment

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    doi: 10.1158/1078-0432.CCR-11-1707

    Attenuated IGF1R and/or EGFR significantly inhibited cell proliferation in MPNST ST88-14 cells by blocking the PI3K/AKT and MAPK pathways
    Figure Legend Snippet: Attenuated IGF1R and/or EGFR significantly inhibited cell proliferation in MPNST ST88-14 cells by blocking the PI3K/AKT and MAPK pathways

    Techniques Used: Blocking Assay

    13) Product Images from "Immobilized epidermal growth factor stimulates persistent, directed keratinocyte migration via activation of PLCγ1"

    Article Title: Immobilized epidermal growth factor stimulates persistent, directed keratinocyte migration via activation of PLCγ1

    Journal: The FASEB Journal

    doi: 10.1096/fj.201600252

    Changes in EGFR localization in response to soluble or immobilized EGF. Immunofluorescent staining for EGFR (green) and EEA1 (red); blue is nuclear stain. Red line indicates the cellular plane shown in the panels. Immob, immobilized; sol, soluble. Scale
    Figure Legend Snippet: Changes in EGFR localization in response to soluble or immobilized EGF. Immunofluorescent staining for EGFR (green) and EEA1 (red); blue is nuclear stain. Red line indicates the cellular plane shown in the panels. Immob, immobilized; sol, soluble. Scale

    Techniques Used: Staining

    14) Product Images from "Epidermal growth factor receptor activity is elevated in glioma cancer stem cells and is required to maintain chemotherapy and radiation resistance"

    Article Title: Epidermal growth factor receptor activity is elevated in glioma cancer stem cells and is required to maintain chemotherapy and radiation resistance

    Journal: Oncotarget

    doi: 10.18632/oncotarget.19868

    EGFR is constitutively active in CSCs (A) J3T adherent and sphere cells were treated with 5 μM gefitinib and harvested over the indicated time course. Expression of EGFR, phosphorylation of EGFR at serine-1047 and tyrosine-1173, AKT, phosphorylation of AKT, and β-actin as a loading control. 20 μg was loaded per lane. Comparison of the effect of increasing doses of gefitinib on proliferation of adherent and spheres of J3T (B) and LN18 cells (C) . Cells were treated with increasing doses of gefitinib and cell viability was assayed 48 hours post treatment.
    Figure Legend Snippet: EGFR is constitutively active in CSCs (A) J3T adherent and sphere cells were treated with 5 μM gefitinib and harvested over the indicated time course. Expression of EGFR, phosphorylation of EGFR at serine-1047 and tyrosine-1173, AKT, phosphorylation of AKT, and β-actin as a loading control. 20 μg was loaded per lane. Comparison of the effect of increasing doses of gefitinib on proliferation of adherent and spheres of J3T (B) and LN18 cells (C) . Cells were treated with increasing doses of gefitinib and cell viability was assayed 48 hours post treatment.

    Techniques Used: Expressing

    15) Product Images from "Integrative Analysis of Membrane Proteome and MicroRNA Reveals Novel Lung Cancer Metastasis Biomarkers"

    Article Title: Integrative Analysis of Membrane Proteome and MicroRNA Reveals Novel Lung Cancer Metastasis Biomarkers

    Journal: Frontiers in Genetics

    doi: 10.3389/fgene.2020.01023

    Kaplan-Meier curves of overall survival for CHE2, EGFR, ITGA3, ITGA5, ITGB1, and CALR.
    Figure Legend Snippet: Kaplan-Meier curves of overall survival for CHE2, EGFR, ITGA3, ITGA5, ITGB1, and CALR.

    Techniques Used:

    Western blot verification results. (A) Western blot assays of the protein level of CDH2, EGFR, ITGA3, TIGB1, TIGA5, and CALR. (B) The gray levels of Western blotting are shown by bar graph.
    Figure Legend Snippet: Western blot verification results. (A) Western blot assays of the protein level of CDH2, EGFR, ITGA3, TIGB1, TIGA5, and CALR. (B) The gray levels of Western blotting are shown by bar graph.

    Techniques Used: Western Blot

    16) Product Images from "CUL4A overexpression enhances lung tumor growth and sensitizes lung cancer cells to Erlotinib via transcriptional regulation of EGFR"

    Article Title: CUL4A overexpression enhances lung tumor growth and sensitizes lung cancer cells to Erlotinib via transcriptional regulation of EGFR

    Journal: Molecular Cancer

    doi: 10.1186/1476-4598-13-252

    CUL4A regulates EGFR expression. (A) RT-PCR analysis of the expression of EGFR mRNA in H1299, H1650, A549 and H460 cells. (B) Western blot analysis of the expression of EGFR protein in H1299, H1650, A549 and H460 cells. (C) Immunofluorescence microscopy analysis of EGFR expression of in H1299, H1650, A549 and H460 cells. (D) The immunohistochemistry analysis of CUL4A and EGFR expression in NSCLC biopsy showed that CUL4A levels significantly correlate with EGFR levels in NSCLC tissues. All results are from three independent experiments. Scale bar indicates 20 μm (C) , and 50 μm (D) .
    Figure Legend Snippet: CUL4A regulates EGFR expression. (A) RT-PCR analysis of the expression of EGFR mRNA in H1299, H1650, A549 and H460 cells. (B) Western blot analysis of the expression of EGFR protein in H1299, H1650, A549 and H460 cells. (C) Immunofluorescence microscopy analysis of EGFR expression of in H1299, H1650, A549 and H460 cells. (D) The immunohistochemistry analysis of CUL4A and EGFR expression in NSCLC biopsy showed that CUL4A levels significantly correlate with EGFR levels in NSCLC tissues. All results are from three independent experiments. Scale bar indicates 20 μm (C) , and 50 μm (D) .

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Immunofluorescence, Microscopy, Immunohistochemistry

    CUL4A transcriptionally activates EGFR expression in NSCLC tissues. (A) Western blot analysis of H3K4me3 levels in H1299-pBabe, H1299-CUL4A, A549-pSuper, and A549-shCUL4A cells. (B) Schematic presentation of two regions relative to the EGFR transcriptional start site used as primers to test H3K4me3 occupied abundance. (C) ChIP-PCR was performed to assess H3K4me3 occupancy in EGFR promoter in H1299-pBabe and H1299-CUL4A cells. (D) ChIP-PCR was performed to assess H3K4me3 occupancy in EGFR promoter in A549-pSuper and A549-shCUL4A cells. IgG was used as negative control.
    Figure Legend Snippet: CUL4A transcriptionally activates EGFR expression in NSCLC tissues. (A) Western blot analysis of H3K4me3 levels in H1299-pBabe, H1299-CUL4A, A549-pSuper, and A549-shCUL4A cells. (B) Schematic presentation of two regions relative to the EGFR transcriptional start site used as primers to test H3K4me3 occupied abundance. (C) ChIP-PCR was performed to assess H3K4me3 occupancy in EGFR promoter in H1299-pBabe and H1299-CUL4A cells. (D) ChIP-PCR was performed to assess H3K4me3 occupancy in EGFR promoter in A549-pSuper and A549-shCUL4A cells. IgG was used as negative control.

    Techniques Used: Expressing, Western Blot, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Negative Control

    CUL4A activates the EGFR-mediated signaling pathways. (A) Levels of CUL4A, EGFR, p-EGFR, p-AKT, and AKT were analyzed by Western blot in H1299-pBabe, H1299-CUL4A, H1650-pBabe and H1650-CUL4A cells. (B) Levels of CUL4A, EGFR, p-EGFR, p-AKT, and AKT were analyzed by Western blot in A549-pSuper, A549-shCUL4A, H460-pSuper and H460-shCUL4A cells. (C) Western blot to analyze the effect of erlotinib on the levels of CUL4A, EGFR, p-EGFR, p-AKT, and AKT in H1299-pBabe and H1299-CUL4A cells. (D) MTT analysis of the inhibition of erlotinib on cell proliferation in CUL4A overexprssion cells (H1299-CUL4A and H1650-CUL4A). ** P
    Figure Legend Snippet: CUL4A activates the EGFR-mediated signaling pathways. (A) Levels of CUL4A, EGFR, p-EGFR, p-AKT, and AKT were analyzed by Western blot in H1299-pBabe, H1299-CUL4A, H1650-pBabe and H1650-CUL4A cells. (B) Levels of CUL4A, EGFR, p-EGFR, p-AKT, and AKT were analyzed by Western blot in A549-pSuper, A549-shCUL4A, H460-pSuper and H460-shCUL4A cells. (C) Western blot to analyze the effect of erlotinib on the levels of CUL4A, EGFR, p-EGFR, p-AKT, and AKT in H1299-pBabe and H1299-CUL4A cells. (D) MTT analysis of the inhibition of erlotinib on cell proliferation in CUL4A overexprssion cells (H1299-CUL4A and H1650-CUL4A). ** P

    Techniques Used: Western Blot, MTT Assay, Inhibition

    17) Product Images from "Dihydrotestosterone Potentiates EGF-Induced ERK Activation by Inducing SRC in Fetal Lung Fibroblasts"

    Article Title: Dihydrotestosterone Potentiates EGF-Induced ERK Activation by Inducing SRC in Fetal Lung Fibroblasts

    Journal: American Journal of Respiratory Cell and Molecular Biology

    doi: 10.1165/rcmb.2012-0179OC

    Src inhibition prevents DHT from enhancing the activity of the EGFR. ( A – C ) PP2 inhibition of SRC prevents DHT from potentiating EGF signaling. Previously untreated primary fetal lung fibroblasts were exposed to PP2 before DHT treatment and EGF
    Figure Legend Snippet: Src inhibition prevents DHT from enhancing the activity of the EGFR. ( A – C ) PP2 inhibition of SRC prevents DHT from potentiating EGF signaling. Previously untreated primary fetal lung fibroblasts were exposed to PP2 before DHT treatment and EGF

    Techniques Used: Inhibition, Activity Assay

    DHT increases SRC phosphorylation and potentiates the ligand-dependent activation of the EGFR. EGF-induced SRC and EGFR pathway activation is potentiated by DHT. Previously untreated primary fetal lung fibroblasts were stimulated with 100 mg/ml EGF for
    Figure Legend Snippet: DHT increases SRC phosphorylation and potentiates the ligand-dependent activation of the EGFR. EGF-induced SRC and EGFR pathway activation is potentiated by DHT. Previously untreated primary fetal lung fibroblasts were stimulated with 100 mg/ml EGF for

    Techniques Used: Activation Assay

    Related Articles

    Western Blot:

    Article Title: Lnc RNA DUXAP9‐206 directly binds with Cbl‐b to augment EGFR signaling and promotes non‐small cell lung cancer progression, et al. LncRNA DUXAP9‐206 directly binds with Cbl‐b to augment EGFR signaling and promotes non‐small cell lung cancer progression
    Article Snippet: After vortex and short centrifugation, the supernatant was collected as cytoplasmic fraction and the remainder with additional washing was considered as nuclear pellets. .. 2.7 Western blotting analysisWestern blotting analysis was performed according to a previously described standard method using the antibodies anti‐Cbl‐b (ab93205, 1:500; Abcam, Cambridge, MA, USA), anti‐EGFR (ab52894, 1:1000; Abcam), anti‐Flag (F2555, 1:2000; Sigma, St Louis, MO, USA), anti‐ubiquitin (ab7780, 1:1000; Abcam), anti‐p‐AKT (ab81283, 1:500; Abcam), anti‐AKT (ab8805, 1:1000; Abcam), anti‐p‐ERK (4370, 1:500; Cell Signaling, Danvers, MA, USA) and anti‐ERK (4695, 1:1000; Cell Signaling). .. The blotted membranes were stripped and re‐blotted with anti‐actin (ab8227, 1:2000; Abcam) as a loading control.

    Article Title: AR–PDEF pathway promotes tumour proliferation and upregulates MYC-mediated gene transcription by promoting MAD1 degradation in ER-negative breast cancer
    Article Snippet: .. Western blotting Western blotting was performed as described previously [ ] by using the following primary antibodies: anti-AR antibody (ab9474; dilution, 1:3000), anti-PDEF antibody (ab53881; dilution, 1:1000; Abcam), anti-MAD1 antibody (ab175245; dilution, 1:3000), anti-MYC antibody (ab32072; dilution, 1:3000), anti-β-catenin antibody (ab32572; dilution, 1:3000; Abcam), anti-AKT antibody (sc-135,829; dilution, 1:3000; Santa Cruz Biotechnology), anti-phosphorylated AKT antibody (anti-p-AKT; sc-7985-R; dilution, 1:3000; Santa Cruz Biotechnology), anti-ERK antibody (sc-514,302; dilution, 1:3000; Santa Cruz Biotechnology), anti-phosphorylated ERK antibody (anti-p-ERK; sc-81,492; dilution, 1:3000; Santa Cruz Biotechnology) and anti-EGFR antibody (ab52894; dilution, 1:3000; Abcam). .. Immunofluorescence staining Immunofluorescence staining was performed as described previously [ ].

    Article Title: Integrating Network Pharmacology and Experimental Validation to Investigate the Mechanisms of Huazhuojiedu Decoction to Treat Chronic Atrophic Gastritis
    Article Snippet: .. Reagents and Equipment Western LightningTM Chemiluminescence Reagent was obtained from PerkinElmer, USA (lot NEL103 001EA); goat anti-rabbit IgG secondary antibody was supplied by Abcam, UK (lot ab6721); rabbit anti-mouse IgG secondary antibody was supplied by Abcam, UK (lot ab6721ab6728); M-mlv reverse transcriptase was obtained from Takara, Japan (lot 2641A); RNase inhibitors were purchased from Takara, Japan (lot D2310 C); SYBR Premix Ex Taq was obtained from Takara, Japan (lot DRR041 A); Hq-350xt development and fixing equipment was supplied by Suzhou Huqiu Image Equipment Co., Ltd., China; Superrx film was supplied by FUJIFILM, Japan; Decolorizing Orbital Shaker Ts-1 was supplied by Jiangsu Haimen Qilin Bell Instrument Manufacturing Co., Ltd., China; Dycz-24dn vertical electrophoresis device was supplied by Beijing Liuyi Instrument Factory, China; Ve-386 transfer electrophoresis tank was supplied by Beijing Yuanpinghao Biotechnology Co., Ltd., China; 5415D centrifuge was supplied by Eppendorf, Germany; Bio-Rad real-time PCR amplification instrument was supplied by Bio-Rad, USA; MAKT1 antibody was obtained from Cell Signaling Technology, USA (cat.4695S); AKT1 antibody was purchased from Abcam, UK (cat. ab81283); TNF antibody was obtained from Abcam, UK (cat. ab6671); VEGFA antibody was supplied by Abcam, UK (cat.ab1316); and EGFR antibody was purchased from Abcam, UK (cat.ab52894). ..

    Immunofluorescence:

    Article Title: EGFR feedback-inhibition by Ran-binding protein 6 is disrupted in cancer
    Article Snippet: DNA fingerprinting was previously performed for authentication of all glioma cell lines . .. Antibodies to RanBP6 (ab74448; 1:1000), EGFR (ab52894; 1:960 for immunofluorescence staining), RanGAP1 (ab92360; 1:200), and CRM1 (ab24189, 1:1000) were purchased from Abcam. .. Antibodies to EGFR (#2085; 1:500 for GST-RanBP6 pulldown and 1:1000–5000 for immunoblot), pEGFR Tyr1068 (#3777; 1:1000), Gab1 (#3232; 1:1000), pGab1 Tyr627 (#3231; 1:1000), pErk1/2 Thr202/Tyr204 (#9101; 1:1000), Akt (#9272S; 1:1000), pAkt Ser473 (#4051; 1:1000), S6 (#2317S; 1:1000), pS6 Ser240/244 (#5364; 1:1000), PTEN (#9556; 1:1000), STAT3 (# 12640S; 1:1000), p-STAT3 Tyr705 (#9145S; 1:1000), Ac-STAT3 Lys685 (#2523S; 1:250), H3 (#4499S; 1:5000), p27Kip1 (#2552, 1:500), RB (#9309, 1:1000), and Foxo3a (#2497, 1:1000) were purchased from Cell Signaling.

    Staining:

    Article Title: EGFR feedback-inhibition by Ran-binding protein 6 is disrupted in cancer
    Article Snippet: DNA fingerprinting was previously performed for authentication of all glioma cell lines . .. Antibodies to RanBP6 (ab74448; 1:1000), EGFR (ab52894; 1:960 for immunofluorescence staining), RanGAP1 (ab92360; 1:200), and CRM1 (ab24189, 1:1000) were purchased from Abcam. .. Antibodies to EGFR (#2085; 1:500 for GST-RanBP6 pulldown and 1:1000–5000 for immunoblot), pEGFR Tyr1068 (#3777; 1:1000), Gab1 (#3232; 1:1000), pGab1 Tyr627 (#3231; 1:1000), pErk1/2 Thr202/Tyr204 (#9101; 1:1000), Akt (#9272S; 1:1000), pAkt Ser473 (#4051; 1:1000), S6 (#2317S; 1:1000), pS6 Ser240/244 (#5364; 1:1000), PTEN (#9556; 1:1000), STAT3 (# 12640S; 1:1000), p-STAT3 Tyr705 (#9145S; 1:1000), Ac-STAT3 Lys685 (#2523S; 1:250), H3 (#4499S; 1:5000), p27Kip1 (#2552, 1:500), RB (#9309, 1:1000), and Foxo3a (#2497, 1:1000) were purchased from Cell Signaling.

    Electrophoresis:

    Article Title: Integrating Network Pharmacology and Experimental Validation to Investigate the Mechanisms of Huazhuojiedu Decoction to Treat Chronic Atrophic Gastritis
    Article Snippet: .. Reagents and Equipment Western LightningTM Chemiluminescence Reagent was obtained from PerkinElmer, USA (lot NEL103 001EA); goat anti-rabbit IgG secondary antibody was supplied by Abcam, UK (lot ab6721); rabbit anti-mouse IgG secondary antibody was supplied by Abcam, UK (lot ab6721ab6728); M-mlv reverse transcriptase was obtained from Takara, Japan (lot 2641A); RNase inhibitors were purchased from Takara, Japan (lot D2310 C); SYBR Premix Ex Taq was obtained from Takara, Japan (lot DRR041 A); Hq-350xt development and fixing equipment was supplied by Suzhou Huqiu Image Equipment Co., Ltd., China; Superrx film was supplied by FUJIFILM, Japan; Decolorizing Orbital Shaker Ts-1 was supplied by Jiangsu Haimen Qilin Bell Instrument Manufacturing Co., Ltd., China; Dycz-24dn vertical electrophoresis device was supplied by Beijing Liuyi Instrument Factory, China; Ve-386 transfer electrophoresis tank was supplied by Beijing Yuanpinghao Biotechnology Co., Ltd., China; 5415D centrifuge was supplied by Eppendorf, Germany; Bio-Rad real-time PCR amplification instrument was supplied by Bio-Rad, USA; MAKT1 antibody was obtained from Cell Signaling Technology, USA (cat.4695S); AKT1 antibody was purchased from Abcam, UK (cat. ab81283); TNF antibody was obtained from Abcam, UK (cat. ab6671); VEGFA antibody was supplied by Abcam, UK (cat.ab1316); and EGFR antibody was purchased from Abcam, UK (cat.ab52894). ..

    Real-time Polymerase Chain Reaction:

    Article Title: Integrating Network Pharmacology and Experimental Validation to Investigate the Mechanisms of Huazhuojiedu Decoction to Treat Chronic Atrophic Gastritis
    Article Snippet: .. Reagents and Equipment Western LightningTM Chemiluminescence Reagent was obtained from PerkinElmer, USA (lot NEL103 001EA); goat anti-rabbit IgG secondary antibody was supplied by Abcam, UK (lot ab6721); rabbit anti-mouse IgG secondary antibody was supplied by Abcam, UK (lot ab6721ab6728); M-mlv reverse transcriptase was obtained from Takara, Japan (lot 2641A); RNase inhibitors were purchased from Takara, Japan (lot D2310 C); SYBR Premix Ex Taq was obtained from Takara, Japan (lot DRR041 A); Hq-350xt development and fixing equipment was supplied by Suzhou Huqiu Image Equipment Co., Ltd., China; Superrx film was supplied by FUJIFILM, Japan; Decolorizing Orbital Shaker Ts-1 was supplied by Jiangsu Haimen Qilin Bell Instrument Manufacturing Co., Ltd., China; Dycz-24dn vertical electrophoresis device was supplied by Beijing Liuyi Instrument Factory, China; Ve-386 transfer electrophoresis tank was supplied by Beijing Yuanpinghao Biotechnology Co., Ltd., China; 5415D centrifuge was supplied by Eppendorf, Germany; Bio-Rad real-time PCR amplification instrument was supplied by Bio-Rad, USA; MAKT1 antibody was obtained from Cell Signaling Technology, USA (cat.4695S); AKT1 antibody was purchased from Abcam, UK (cat. ab81283); TNF antibody was obtained from Abcam, UK (cat. ab6671); VEGFA antibody was supplied by Abcam, UK (cat.ab1316); and EGFR antibody was purchased from Abcam, UK (cat.ab52894). ..

    Amplification:

    Article Title: Integrating Network Pharmacology and Experimental Validation to Investigate the Mechanisms of Huazhuojiedu Decoction to Treat Chronic Atrophic Gastritis
    Article Snippet: .. Reagents and Equipment Western LightningTM Chemiluminescence Reagent was obtained from PerkinElmer, USA (lot NEL103 001EA); goat anti-rabbit IgG secondary antibody was supplied by Abcam, UK (lot ab6721); rabbit anti-mouse IgG secondary antibody was supplied by Abcam, UK (lot ab6721ab6728); M-mlv reverse transcriptase was obtained from Takara, Japan (lot 2641A); RNase inhibitors were purchased from Takara, Japan (lot D2310 C); SYBR Premix Ex Taq was obtained from Takara, Japan (lot DRR041 A); Hq-350xt development and fixing equipment was supplied by Suzhou Huqiu Image Equipment Co., Ltd., China; Superrx film was supplied by FUJIFILM, Japan; Decolorizing Orbital Shaker Ts-1 was supplied by Jiangsu Haimen Qilin Bell Instrument Manufacturing Co., Ltd., China; Dycz-24dn vertical electrophoresis device was supplied by Beijing Liuyi Instrument Factory, China; Ve-386 transfer electrophoresis tank was supplied by Beijing Yuanpinghao Biotechnology Co., Ltd., China; 5415D centrifuge was supplied by Eppendorf, Germany; Bio-Rad real-time PCR amplification instrument was supplied by Bio-Rad, USA; MAKT1 antibody was obtained from Cell Signaling Technology, USA (cat.4695S); AKT1 antibody was purchased from Abcam, UK (cat. ab81283); TNF antibody was obtained from Abcam, UK (cat. ab6671); VEGFA antibody was supplied by Abcam, UK (cat.ab1316); and EGFR antibody was purchased from Abcam, UK (cat.ab52894). ..

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  • 94
    Abcam anti cd133
    ABCG1 regulates proliferation-, apoptosis- and cancer stem cell-associated markers in HKULC4 lung cancer cells. Western blot for c-Myc, p21, p53, Ki67, RB, BCL2, MCL1, <t>CD133</t> and ALDH1 in HKULC4 cells transfected with ABCG1-expressing plasmid or empty vector. β-actin was used as a loading control (representative images of three experiments are shown). ABCG1, ATP-binding cassette transporter G1; BCL2, B-cell lymphoma 2; MCL1, myeloid cell leukemia 1; ALDH, aldehyde dehydrogenase; RB, retinoblastoma protein.
    Anti Cd133, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    egfr  (Abcam)
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    Abcam egfr
    Effect of gefitinib on cell proliferation and targeted markers in pancreatic tumors. Immunohistochemical analysis of proliferating cell nuclear antigen (PCNA) (A), β-catanin (B); <t>EGFR</t> (C), and Caveolin-1 (D) expressions in normal pancreatic tissues and ductal adenocarcinomas. Immunoblotting was performed with paraffin embed and micro-sectioned pancreatic tissues as described in the methods section. Fig 3E, Effect of gefitinib on the inhibition of PanINs thereby mucin expression (blue color) in pancreatic tissues by alcin blue staining method.
    Egfr, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pegfr  (Abcam)
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    Abcam pegfr
    Cell surface molecules regulated by mutant p53 are involved in CIC formation. a 3D images of a CIC structure in H1299 cells. The top right insert is showing the horizontal plane by confocal imaging. Scale bar = 5 μm. b Fluorescent image of live A431 cells that have been labeled with pHrodo. Scale bars = 50 μm. c Confocal image of A431 cells that have been stained with Calcein AM. Scale bars = 20 μm. d Quantification of CIC for H1299 co-cultures seeded on plastics coated with different matrices. Each bar represents +/−SEM of triplicate experiments ** p = 0.0072. e Immunofluorescence image of a CIC structure formed between H1299 cells (green and red as indicated). Dapi is in blue and CD49e (integrin α5) is in yellow. Scale bar = 50 μm. f Number of CIC events in mutant p53 (green)/KO p53 (red) A431 co-cultures in which engulfment by red cells was quantified in the presence of mAb16. Each bar represents +/−SEM of triplicate experiments ** p = 0.003. g Quantification of CIC in H1299 EV/H1299 175H co-cultures in the presence or absence of Y27632 or mAb16. Each bar represents +/−SEM of triplicate experiments * p = 0.01–0.04. h Quantification of total CIC structures per 20 hpf observed in A431 Ctr 273H/mCherry and KO 273H/GFP co-cultures treated with and without EGF for 24 h. Either red or green engulfing cells were quantified for KO 273H or ctr 273H respectively. Each bar represents +/−SEM of triplicate experiments Each bar represents +/−SEM of triplicate experiments * p = 0.03 or ** p = 0.004. i Quantification of total CIC structures per 20 hpf observed in A431 Ctr 273H/mCherry and KO 273H/GFP co-cultured cells treated with and without Gefitinib (24 h). Either red or green engulfing cells were quantified for KO 273H or ctr 273H respectively Each bar represents +/−SEM of triplicate experiments ** p = 0.0019. j Western blot showing <t>pEGFR</t> and <t>EGFR</t> following treatment of A431 control and p53 CRISPR KO cells, with and without Gefitinib and EGF. Actin was used as loading control
    Pegfr, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti egfr
    Association of Cell surface GRP78 with α2M* facilitated the maximal activation of <t>EGFR</t> in a c-Src dependent manner. (a) Western blot analysis of the expression and phosphorylation of EGFR in serum starved QGY-7703 treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*. (b) Quantitative analysis of the expression and phosphorylation of EGFR in serum starved QGY-7703 treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*. (c) <t>Co-immunoprecipitation</t> analysis of the interaction between c-Src and EGFR in serum starved QGY-7703 cells treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*. (d) Quantitative analysis of the interaction between c-Src and EGFR in serum starved QGY-7703 cells treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*
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    Image Search Results


    ABCG1 regulates proliferation-, apoptosis- and cancer stem cell-associated markers in HKULC4 lung cancer cells. Western blot for c-Myc, p21, p53, Ki67, RB, BCL2, MCL1, CD133 and ALDH1 in HKULC4 cells transfected with ABCG1-expressing plasmid or empty vector. β-actin was used as a loading control (representative images of three experiments are shown). ABCG1, ATP-binding cassette transporter G1; BCL2, B-cell lymphoma 2; MCL1, myeloid cell leukemia 1; ALDH, aldehyde dehydrogenase; RB, retinoblastoma protein.

    Journal: Experimental and Therapeutic Medicine

    Article Title: ABCG1 as a potential oncogene in lung cancer

    doi: 10.3892/etm.2017.4393

    Figure Lengend Snippet: ABCG1 regulates proliferation-, apoptosis- and cancer stem cell-associated markers in HKULC4 lung cancer cells. Western blot for c-Myc, p21, p53, Ki67, RB, BCL2, MCL1, CD133 and ALDH1 in HKULC4 cells transfected with ABCG1-expressing plasmid or empty vector. β-actin was used as a loading control (representative images of three experiments are shown). ABCG1, ATP-binding cassette transporter G1; BCL2, B-cell lymphoma 2; MCL1, myeloid cell leukemia 1; ALDH, aldehyde dehydrogenase; RB, retinoblastoma protein.

    Article Snippet: Membranes were blocked with 5% skimmed milk and incubated overnight at 4°C with primary antibodies, including rabbit anti-human monoclonal anti-ABCG1 (1:5,000; ab52617), anti-c-Myc (1:5,000; ab32072), anti-p21 (1:5,000; ab109520), anti-p53 (1:5,000; ab32049), anti-Ki-67 (1:5,000; ab15580), anti-retinoblastoma (1:5,000; ab47763), anti-B-cell lymphoma 2 (BCL2; 1:5,000; ab23124), anti-myeloid cell leukemia 1 (MCL1; 1:5,000; ab32087), anti-CD133 (1:5,000; ab32077) and anti-aldehyde dehydrogenase (ALDH; 1:5,000; ab52492; all Abcam, Cambridge, MA, USA).

    Techniques: Western Blot, Transfection, Expressing, Plasmid Preparation, Binding Assay

    Effect of gefitinib on cell proliferation and targeted markers in pancreatic tumors. Immunohistochemical analysis of proliferating cell nuclear antigen (PCNA) (A), β-catanin (B); EGFR (C), and Caveolin-1 (D) expressions in normal pancreatic tissues and ductal adenocarcinomas. Immunoblotting was performed with paraffin embed and micro-sectioned pancreatic tissues as described in the methods section. Fig 3E, Effect of gefitinib on the inhibition of PanINs thereby mucin expression (blue color) in pancreatic tissues by alcin blue staining method.

    Journal: Cancer prevention research (Philadelphia, Pa.)

    Article Title: EGFR inhibitor gefitinib prevents progression of pancreatic lesions to carcinoma in a conditional LSL-KrasG12D/+ transgenic mice model

    doi: 10.1158/1940-6207.CAPR-10-0038

    Figure Lengend Snippet: Effect of gefitinib on cell proliferation and targeted markers in pancreatic tumors. Immunohistochemical analysis of proliferating cell nuclear antigen (PCNA) (A), β-catanin (B); EGFR (C), and Caveolin-1 (D) expressions in normal pancreatic tissues and ductal adenocarcinomas. Immunoblotting was performed with paraffin embed and micro-sectioned pancreatic tissues as described in the methods section. Fig 3E, Effect of gefitinib on the inhibition of PanINs thereby mucin expression (blue color) in pancreatic tissues by alcin blue staining method.

    Article Snippet: Sections were then incubated overnight at 4°C with 1:300 dilutions of monoclonal antibodies against PCNA, β-catenin, EGFR and Cav-1 (AbCam/Santa Cruz Biotechnology, CA).

    Techniques: Immunohistochemistry, Inhibition, Expressing, Staining

    Cell surface molecules regulated by mutant p53 are involved in CIC formation. a 3D images of a CIC structure in H1299 cells. The top right insert is showing the horizontal plane by confocal imaging. Scale bar = 5 μm. b Fluorescent image of live A431 cells that have been labeled with pHrodo. Scale bars = 50 μm. c Confocal image of A431 cells that have been stained with Calcein AM. Scale bars = 20 μm. d Quantification of CIC for H1299 co-cultures seeded on plastics coated with different matrices. Each bar represents +/−SEM of triplicate experiments ** p = 0.0072. e Immunofluorescence image of a CIC structure formed between H1299 cells (green and red as indicated). Dapi is in blue and CD49e (integrin α5) is in yellow. Scale bar = 50 μm. f Number of CIC events in mutant p53 (green)/KO p53 (red) A431 co-cultures in which engulfment by red cells was quantified in the presence of mAb16. Each bar represents +/−SEM of triplicate experiments ** p = 0.003. g Quantification of CIC in H1299 EV/H1299 175H co-cultures in the presence or absence of Y27632 or mAb16. Each bar represents +/−SEM of triplicate experiments * p = 0.01–0.04. h Quantification of total CIC structures per 20 hpf observed in A431 Ctr 273H/mCherry and KO 273H/GFP co-cultures treated with and without EGF for 24 h. Either red or green engulfing cells were quantified for KO 273H or ctr 273H respectively. Each bar represents +/−SEM of triplicate experiments Each bar represents +/−SEM of triplicate experiments * p = 0.03 or ** p = 0.004. i Quantification of total CIC structures per 20 hpf observed in A431 Ctr 273H/mCherry and KO 273H/GFP co-cultured cells treated with and without Gefitinib (24 h). Either red or green engulfing cells were quantified for KO 273H or ctr 273H respectively Each bar represents +/−SEM of triplicate experiments ** p = 0.0019. j Western blot showing pEGFR and EGFR following treatment of A431 control and p53 CRISPR KO cells, with and without Gefitinib and EGF. Actin was used as loading control

    Journal: Nature Communications

    Article Title: Genomic instability in mutant p53 cancer cells upon entotic engulfment

    doi: 10.1038/s41467-018-05368-1

    Figure Lengend Snippet: Cell surface molecules regulated by mutant p53 are involved in CIC formation. a 3D images of a CIC structure in H1299 cells. The top right insert is showing the horizontal plane by confocal imaging. Scale bar = 5 μm. b Fluorescent image of live A431 cells that have been labeled with pHrodo. Scale bars = 50 μm. c Confocal image of A431 cells that have been stained with Calcein AM. Scale bars = 20 μm. d Quantification of CIC for H1299 co-cultures seeded on plastics coated with different matrices. Each bar represents +/−SEM of triplicate experiments ** p = 0.0072. e Immunofluorescence image of a CIC structure formed between H1299 cells (green and red as indicated). Dapi is in blue and CD49e (integrin α5) is in yellow. Scale bar = 50 μm. f Number of CIC events in mutant p53 (green)/KO p53 (red) A431 co-cultures in which engulfment by red cells was quantified in the presence of mAb16. Each bar represents +/−SEM of triplicate experiments ** p = 0.003. g Quantification of CIC in H1299 EV/H1299 175H co-cultures in the presence or absence of Y27632 or mAb16. Each bar represents +/−SEM of triplicate experiments * p = 0.01–0.04. h Quantification of total CIC structures per 20 hpf observed in A431 Ctr 273H/mCherry and KO 273H/GFP co-cultures treated with and without EGF for 24 h. Either red or green engulfing cells were quantified for KO 273H or ctr 273H respectively. Each bar represents +/−SEM of triplicate experiments Each bar represents +/−SEM of triplicate experiments * p = 0.03 or ** p = 0.004. i Quantification of total CIC structures per 20 hpf observed in A431 Ctr 273H/mCherry and KO 273H/GFP co-cultured cells treated with and without Gefitinib (24 h). Either red or green engulfing cells were quantified for KO 273H or ctr 273H respectively Each bar represents +/−SEM of triplicate experiments ** p = 0.0019. j Western blot showing pEGFR and EGFR following treatment of A431 control and p53 CRISPR KO cells, with and without Gefitinib and EGF. Actin was used as loading control

    Article Snippet: The following primary antibodies were used for western blotting: EGFR (Cell Signaling), pEGFR (Abcam), Actin (Millipore).The blots were washed three times with TBS-T and then incubated with corresponding IRDye secondary antibodies 680RD or 800CW (Li-Cor, Cambridge, UK, 0.05 µg/ml).

    Techniques: Mutagenesis, Imaging, Labeling, Staining, Immunofluorescence, Cell Culture, Western Blot, CRISPR

    Association of Cell surface GRP78 with α2M* facilitated the maximal activation of EGFR in a c-Src dependent manner. (a) Western blot analysis of the expression and phosphorylation of EGFR in serum starved QGY-7703 treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*. (b) Quantitative analysis of the expression and phosphorylation of EGFR in serum starved QGY-7703 treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*. (c) Co-immunoprecipitation analysis of the interaction between c-Src and EGFR in serum starved QGY-7703 cells treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*. (d) Quantitative analysis of the interaction between c-Src and EGFR in serum starved QGY-7703 cells treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*

    Journal: BMC Cancer

    Article Title: The role of c-Src in the invasion and metastasis of hepatocellular carcinoma cells induced by association of cell surface GRP78 with activated α2M

    doi: 10.1186/s12885-015-1401-z

    Figure Lengend Snippet: Association of Cell surface GRP78 with α2M* facilitated the maximal activation of EGFR in a c-Src dependent manner. (a) Western blot analysis of the expression and phosphorylation of EGFR in serum starved QGY-7703 treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*. (b) Quantitative analysis of the expression and phosphorylation of EGFR in serum starved QGY-7703 treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*. (c) Co-immunoprecipitation analysis of the interaction between c-Src and EGFR in serum starved QGY-7703 cells treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*. (d) Quantitative analysis of the interaction between c-Src and EGFR in serum starved QGY-7703 cells treated with vehicle, α2M*, PP2 or PP2 in combination with α2M*

    Article Snippet: Anti-GRP78 (for immunoprecipitation and in cell western analysis), anti-pEGFR Y1101, anti-pEGFR Y1068, anti-pEGFR Y845, anti-EGFR, anti-p-Tyr and rabbit isotype IgG were obtained from Abcam.

    Techniques: Activation Assay, Western Blot, Expressing, Immunoprecipitation