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  • 99
    Name:
    Epidermal Growth Factor Human EGF
    Description:
    The human EGF coding cDNA was obtained from human periodontal tissue mRNA subcloned into a prokaryotic expression vector and expressed in E coli Epidermal Growth Factor Human EGF was purified and stored in PBS buffer containing 0 1 BSA
    Catalog Number:
    9908
    Price:
    None
    Category:
    Cytokines
    Source:
    Human Recombinant Protein
    Buy from Supplier


    Structured Review

    Cell Signaling Technology Inc p egfr
    Differential expression of <t>EGFR,</t> p-EGFR and <t>PKC-α</t> between MEKi-Se and MEKi-Re LGSC cell lines by WB. a Confirmation of the RPPA results in untreated MEKi-Se and MEKi-Re lines ( a , discovery cohort). EGFR, p-EGFR and PKC-α were increased in MEKi-Re lines (n = 5) compared to MEKi-Se lines (n = 2). b Validation of these protein biomarkers in three newly established LGSC cell lines classified according to their MEKi responsiveness (validation cohort). As found in the cell lines analyzed by RPPA, the new MEKi-Re line (n = 1) expressed higher levels of EGFR, p-EGFR and PKC-α compared to the two new MEKi-Se lines tested (n = 2). c , d Confirmation of RPPA results in MEKi treated cell lines. With MEKi treatment (selumetinib 1 μM and trametinib 0.1 μM) p-EGFR expression remained higher in MEKi-Re lines. As previously described, trametinib showed stronger inhibitory effects on MAPK (p-MAPK or p-ERK1/2) than selumetinib, even when used at ten times lower dose. (*) No DMSO. (**) Untreated VOA-4627 cells to control for drug inhibition effects on MAPK pathway
    The human EGF coding cDNA was obtained from human periodontal tissue mRNA subcloned into a prokaryotic expression vector and expressed in E coli Epidermal Growth Factor Human EGF was purified and stored in PBS buffer containing 0 1 BSA
    https://www.bioz.com/result/p egfr/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p egfr - by Bioz Stars, 2021-05
    99/100 stars

    Images

    1) Product Images from "Markers of MEK inhibitor resistance in low-grade serous ovarian cancer: EGFR is a potential therapeutic target"

    Article Title: Markers of MEK inhibitor resistance in low-grade serous ovarian cancer: EGFR is a potential therapeutic target

    Journal: Cancer Cell International

    doi: 10.1186/s12935-019-0725-1

    Differential expression of EGFR, p-EGFR and PKC-α between MEKi-Se and MEKi-Re LGSC cell lines by WB. a Confirmation of the RPPA results in untreated MEKi-Se and MEKi-Re lines ( a , discovery cohort). EGFR, p-EGFR and PKC-α were increased in MEKi-Re lines (n = 5) compared to MEKi-Se lines (n = 2). b Validation of these protein biomarkers in three newly established LGSC cell lines classified according to their MEKi responsiveness (validation cohort). As found in the cell lines analyzed by RPPA, the new MEKi-Re line (n = 1) expressed higher levels of EGFR, p-EGFR and PKC-α compared to the two new MEKi-Se lines tested (n = 2). c , d Confirmation of RPPA results in MEKi treated cell lines. With MEKi treatment (selumetinib 1 μM and trametinib 0.1 μM) p-EGFR expression remained higher in MEKi-Re lines. As previously described, trametinib showed stronger inhibitory effects on MAPK (p-MAPK or p-ERK1/2) than selumetinib, even when used at ten times lower dose. (*) No DMSO. (**) Untreated VOA-4627 cells to control for drug inhibition effects on MAPK pathway
    Figure Legend Snippet: Differential expression of EGFR, p-EGFR and PKC-α between MEKi-Se and MEKi-Re LGSC cell lines by WB. a Confirmation of the RPPA results in untreated MEKi-Se and MEKi-Re lines ( a , discovery cohort). EGFR, p-EGFR and PKC-α were increased in MEKi-Re lines (n = 5) compared to MEKi-Se lines (n = 2). b Validation of these protein biomarkers in three newly established LGSC cell lines classified according to their MEKi responsiveness (validation cohort). As found in the cell lines analyzed by RPPA, the new MEKi-Re line (n = 1) expressed higher levels of EGFR, p-EGFR and PKC-α compared to the two new MEKi-Se lines tested (n = 2). c , d Confirmation of RPPA results in MEKi treated cell lines. With MEKi treatment (selumetinib 1 μM and trametinib 0.1 μM) p-EGFR expression remained higher in MEKi-Re lines. As previously described, trametinib showed stronger inhibitory effects on MAPK (p-MAPK or p-ERK1/2) than selumetinib, even when used at ten times lower dose. (*) No DMSO. (**) Untreated VOA-4627 cells to control for drug inhibition effects on MAPK pathway

    Techniques Used: Expressing, Western Blot, Inhibition

    2) Product Images from "Cx32 exerts anti-apoptotic and pro-tumor effects via the epidermal growth factor receptor pathway in hepatocellular carcinoma"

    Article Title: Cx32 exerts anti-apoptotic and pro-tumor effects via the epidermal growth factor receptor pathway in hepatocellular carcinoma

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/s13046-019-1142-y

    Overexpression of Cx32 promotes the proliferation of SMMC-7721 cells and protects cells from SN-induced apoptosis in vivo. a. Representative images of the nude mouse xenograft model. b. Tumor growth curves. Overexpression of Cx32 promoted the tumor growth in nude mice and significantly reduced the growth suppression mediated by intragastric injection of SN (0.5 mg/kg). c. Representative images of tumors from the sacrificed nude mice. d. Representative images of IHC for Cx32, EGFR and Src in tumors generated from SMMC-Vector and SMMC-Cx32 cells. Scale bars: 50 μm. e. Overexpression of Cx32 inhibited the SN-induced increase in the levels of cleaved-caspase3 by increasing the levels of EGFR and Src. ## , P
    Figure Legend Snippet: Overexpression of Cx32 promotes the proliferation of SMMC-7721 cells and protects cells from SN-induced apoptosis in vivo. a. Representative images of the nude mouse xenograft model. b. Tumor growth curves. Overexpression of Cx32 promoted the tumor growth in nude mice and significantly reduced the growth suppression mediated by intragastric injection of SN (0.5 mg/kg). c. Representative images of tumors from the sacrificed nude mice. d. Representative images of IHC for Cx32, EGFR and Src in tumors generated from SMMC-Vector and SMMC-Cx32 cells. Scale bars: 50 μm. e. Overexpression of Cx32 inhibited the SN-induced increase in the levels of cleaved-caspase3 by increasing the levels of EGFR and Src. ## , P

    Techniques Used: Over Expression, In Vivo, Mouse Assay, Injection, Immunohistochemistry, Generated, Plasmid Preparation

    3) Product Images from "Exposure of Barrett’s and esophageal adenocarcinoma cells to Bile acids activates EGFR-STAT3 signaling axis via induction of APE1"

    Article Title: Exposure of Barrett’s and esophageal adenocarcinoma cells to Bile acids activates EGFR-STAT3 signaling axis via induction of APE1

    Journal: Oncogene

    doi: 10.1038/s41388-018-0388-8

    APE1 mediates bile salts-induced STAT3 activation via an EGFR-dependent mechanism Immunoblot analysis of CPB ( A ), FLO-1 ( B ) and OE33 ( C ) cells pretreated with EGFR inhibitor (Gefitinib, 25 μM) followed by exposure to acidic (pH4) bile salts (100 μM). The samples were analyzed for the indicated proteins, β-actin was used as an internal control. ( D ) Immunoblot analysis of OE33 cells with EGFR-knockdown via EGFR siRNA followed by treatment with acidic (pH4) bile salts (100 μM) for 20 minutes and allowed to recover in complete media. The samples were collected at 3 and 6h post recovery and analyzed for the indicated proteins, β-actin was used as an internal control. Immunoprecipitation (IP) of APE1 ( E ), EGFR ( F ) and STAT3 ( G ) in OE33 cells treated with acidic (pH4) bile salts (100 μM) and immunoblotted for the indicated proteins. Results shown are representative of at least three independent experiments.
    Figure Legend Snippet: APE1 mediates bile salts-induced STAT3 activation via an EGFR-dependent mechanism Immunoblot analysis of CPB ( A ), FLO-1 ( B ) and OE33 ( C ) cells pretreated with EGFR inhibitor (Gefitinib, 25 μM) followed by exposure to acidic (pH4) bile salts (100 μM). The samples were analyzed for the indicated proteins, β-actin was used as an internal control. ( D ) Immunoblot analysis of OE33 cells with EGFR-knockdown via EGFR siRNA followed by treatment with acidic (pH4) bile salts (100 μM) for 20 minutes and allowed to recover in complete media. The samples were collected at 3 and 6h post recovery and analyzed for the indicated proteins, β-actin was used as an internal control. Immunoprecipitation (IP) of APE1 ( E ), EGFR ( F ) and STAT3 ( G ) in OE33 cells treated with acidic (pH4) bile salts (100 μM) and immunoblotted for the indicated proteins. Results shown are representative of at least three independent experiments.

    Techniques Used: Activation Assay, Immunoprecipitation

    APE1 co-localizes with EGFR and STAT3 (A and B) In situ proximity ligation assay (PLA) demonstrates the interaction of p-EGFR and p-STAT3 with APE1. (C) In situ proximity ligation assay (PLA) demonstrates the interaction of p-EGFR and p-STAT3. Protein interactions (red fluorescent signals) were revealed by PLA anti-rabbit plus probe and PLA anti-mouse minus probe in OE33 cells treated with acidic (pH4) bile salts (100 μM) as mentioned in the materials and methods. Nuclei were stained with DAPI (blue). Results shown are representative of at least three independent experiments.
    Figure Legend Snippet: APE1 co-localizes with EGFR and STAT3 (A and B) In situ proximity ligation assay (PLA) demonstrates the interaction of p-EGFR and p-STAT3 with APE1. (C) In situ proximity ligation assay (PLA) demonstrates the interaction of p-EGFR and p-STAT3. Protein interactions (red fluorescent signals) were revealed by PLA anti-rabbit plus probe and PLA anti-mouse minus probe in OE33 cells treated with acidic (pH4) bile salts (100 μM) as mentioned in the materials and methods. Nuclei were stained with DAPI (blue). Results shown are representative of at least three independent experiments.

    Techniques Used: In Situ, Proximity Ligation Assay, Staining

    4) Product Images from "Elevated Expression of Fn14 in Non-Small Cell Lung Cancer Correlates with Activated EGFR and Promotes Tumor Cell Migration and Invasion"

    Article Title: Elevated Expression of Fn14 in Non-Small Cell Lung Cancer Correlates with Activated EGFR and Promotes Tumor Cell Migration and Invasion

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2012.03.026

    Ectopic expression of wild-type (WT) EGFR, activated EGFR mutants, and the K-ras V12 mutant in rat lung epithelial cells (RL-65) induces Fn14 expression. Total cellular lysates from RL-65 cells stably expressing the indicated EGFR receptors or K-ras V12
    Figure Legend Snippet: Ectopic expression of wild-type (WT) EGFR, activated EGFR mutants, and the K-ras V12 mutant in rat lung epithelial cells (RL-65) induces Fn14 expression. Total cellular lysates from RL-65 cells stably expressing the indicated EGFR receptors or K-ras V12

    Techniques Used: Expressing, Mutagenesis, Stable Transfection

    Erlotinib treatment of TKI-sensitive NSCLC cells decreases Fn14 expression. A: Serum-starved HCC827 cells containing the EGFR E746-A750–activating mutation were treated with vehicle for 12 hours or 1 μmol/L erlotinib for the indicated
    Figure Legend Snippet: Erlotinib treatment of TKI-sensitive NSCLC cells decreases Fn14 expression. A: Serum-starved HCC827 cells containing the EGFR E746-A750–activating mutation were treated with vehicle for 12 hours or 1 μmol/L erlotinib for the indicated

    Techniques Used: Expressing, Mutagenesis

    Fn14 expression in human NSCLC specimens and correlation with EGFR phosphorylation. A: Fn14 and p-EGFR staining on representative samples from two patients with lung adenocarcinoma (×5 objective, Aperio GL Scanner; Aperio, Vista, CA). Insets:
    Figure Legend Snippet: Fn14 expression in human NSCLC specimens and correlation with EGFR phosphorylation. A: Fn14 and p-EGFR staining on representative samples from two patients with lung adenocarcinoma (×5 objective, Aperio GL Scanner; Aperio, Vista, CA). Insets:

    Techniques Used: Expressing, Staining

    5) Product Images from "Inhibition of histone deacetylases sensitizes EGF receptor‐TK inhibitor‐resistant non‐small‐cell lung cancer cells to erlotinib in vitro and in vivo) Inhibition of histone deacetylases sensitizes EGF receptor‐TK inhibitor‐resistant non‐small‐cell lung cancer cells to erlotinib in vitro and in vivo"

    Article Title: Inhibition of histone deacetylases sensitizes EGF receptor‐TK inhibitor‐resistant non‐small‐cell lung cancer cells to erlotinib in vitro and in vivo) Inhibition of histone deacetylases sensitizes EGF receptor‐TK inhibitor‐resistant non‐small‐cell lung cancer cells to erlotinib in vitro and in vivo

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.13961

    YF454A plus erlotinib significantly suppresses the receptor tyrosine kinase signalling pathways in EGFR‐TKI‐resistant NSCLC cells. (A) Effects of either YF454A or erlotinib alone or in combination on the protein and mRNA expression of EGFR, Her2, AXL, c‐Met and IGF‐1R. PC9/ER cells were treated with YF454A (0.2 μM), erlotinib (5 μM) or YF454A/erlotinib for 24 h. Cell lysates and total mRNA were prepared, and Western blotting assays ( i ) and real‐time PCR analysis ( ii ) were further carried out. (B) The combination of YF454A and erlotinib time‐dependently suppressed protein expression ( i ) and mRNA expression ( ii ) of EGFR, HER2, AXL, MET and IGF‐1R. (C, D) YF454A/erlotinib down‐regulated the phosphorylation of Akt and ERK in PC9/ER cells (C) and in HCC827/ER cells (D). PC9/ER and HCC827/ER cells were treated with YF454A (0.2 μM) and erlotinib (5 μM) as a single agent alone or in combination for 24 h. The whole‐cell lysates were prepared and probed with specific Akt and ERK antibodies. Western blotting and real‐time PCR assays were performed three independent times, and representative images are shown.
    Figure Legend Snippet: YF454A plus erlotinib significantly suppresses the receptor tyrosine kinase signalling pathways in EGFR‐TKI‐resistant NSCLC cells. (A) Effects of either YF454A or erlotinib alone or in combination on the protein and mRNA expression of EGFR, Her2, AXL, c‐Met and IGF‐1R. PC9/ER cells were treated with YF454A (0.2 μM), erlotinib (5 μM) or YF454A/erlotinib for 24 h. Cell lysates and total mRNA were prepared, and Western blotting assays ( i ) and real‐time PCR analysis ( ii ) were further carried out. (B) The combination of YF454A and erlotinib time‐dependently suppressed protein expression ( i ) and mRNA expression ( ii ) of EGFR, HER2, AXL, MET and IGF‐1R. (C, D) YF454A/erlotinib down‐regulated the phosphorylation of Akt and ERK in PC9/ER cells (C) and in HCC827/ER cells (D). PC9/ER and HCC827/ER cells were treated with YF454A (0.2 μM) and erlotinib (5 μM) as a single agent alone or in combination for 24 h. The whole‐cell lysates were prepared and probed with specific Akt and ERK antibodies. Western blotting and real‐time PCR assays were performed three independent times, and representative images are shown.

    Techniques Used: Expressing, Western Blot, Real-time Polymerase Chain Reaction

    Molecular characterization of EGFR‐TKI‐resistant NSCLC cells. (A) Cytotoxicity of erlotinib in PC9, PC9/ER, HCC827 and HCC827/ER cells was determined using the cell viability assays. Means from three independent experiments are shown with bars representing SEM ( n = 3 per group). (B) Characterization of phosphorylation status of tyrosine kinases including Her2, EGFR, Met, IGF1R, AXL and their downstream molecules Akt and ERK in PC9, PC9/ER, HCC827 and HCC827/ER cells respectively.
    Figure Legend Snippet: Molecular characterization of EGFR‐TKI‐resistant NSCLC cells. (A) Cytotoxicity of erlotinib in PC9, PC9/ER, HCC827 and HCC827/ER cells was determined using the cell viability assays. Means from three independent experiments are shown with bars representing SEM ( n = 3 per group). (B) Characterization of phosphorylation status of tyrosine kinases including Her2, EGFR, Met, IGF1R, AXL and their downstream molecules Akt and ERK in PC9, PC9/ER, HCC827 and HCC827/ER cells respectively.

    Techniques Used:

    6) Product Images from "EREG-driven oncogenesis of Head and Neck Squamous Cell Carcinoma exhibits higher sensitivity to Erlotinib therapy"

    Article Title: EREG-driven oncogenesis of Head and Neck Squamous Cell Carcinoma exhibits higher sensitivity to Erlotinib therapy

    Journal: Theranostics

    doi: 10.7150/thno.47176

    EREG binds to EGFR via N57 and requires the EGFR domains I and III. (A) HEK293 cells were transiently coexpressed with FLAG-EREG and HA-tagged EGFR. Cell extracts were immunoprecipitated separately with anti-FLAG or anti-HA antibodies, and the associated EGFR and EREG proteins were examined by Western blotting. (B) Endogenous EGFR and EREG were immunoprecipitated from HN4 cells, and bound endogenous EREG and EGFR were examined by Western blotting. (C) The cellular location of EGFR (red) and EREG (green) was examined by immunofluorescence staining (nuclei were stained with DAPI; blue). Scale bar, 50 μm. (D) Schematic diagram of the WT, domain I deletion (∆D1), and domain III mutation (D355T/F357A) constructs of EGFR (FLAG-EGFR-ECD). The numbers represent amino acid residues. (E-F) FLAG-tagged WT or deletion mutants of EGFR were coexpressed with HA-EREG in HEK293 cells. Extracts were immunoprecipitated with an anti-FLAG or anti-HA antibody, and bound EREG or EGFR was examined by Western blotting using the anti-HA or anti-FLAG antibody (for input control, see Figure S4C ). (G) Sequence alignment of EREG from different species. (H) Schematic diagram of various EREG NQ mutants used in this study. The numbers indicate amino acid positions on the EREG. (I-J) HA-tagged WT or NQ mutants of EREG were coexpressed with FLAG-EGFR in HEK293 cells. EREG and EGFR were immunoprecipitated with anti-HA and anti-FLAG antibodies, respectively, and analyzed by Western blotting.
    Figure Legend Snippet: EREG binds to EGFR via N57 and requires the EGFR domains I and III. (A) HEK293 cells were transiently coexpressed with FLAG-EREG and HA-tagged EGFR. Cell extracts were immunoprecipitated separately with anti-FLAG or anti-HA antibodies, and the associated EGFR and EREG proteins were examined by Western blotting. (B) Endogenous EGFR and EREG were immunoprecipitated from HN4 cells, and bound endogenous EREG and EGFR were examined by Western blotting. (C) The cellular location of EGFR (red) and EREG (green) was examined by immunofluorescence staining (nuclei were stained with DAPI; blue). Scale bar, 50 μm. (D) Schematic diagram of the WT, domain I deletion (∆D1), and domain III mutation (D355T/F357A) constructs of EGFR (FLAG-EGFR-ECD). The numbers represent amino acid residues. (E-F) FLAG-tagged WT or deletion mutants of EGFR were coexpressed with HA-EREG in HEK293 cells. Extracts were immunoprecipitated with an anti-FLAG or anti-HA antibody, and bound EREG or EGFR was examined by Western blotting using the anti-HA or anti-FLAG antibody (for input control, see Figure S4C ). (G) Sequence alignment of EREG from different species. (H) Schematic diagram of various EREG NQ mutants used in this study. The numbers indicate amino acid positions on the EREG. (I-J) HA-tagged WT or NQ mutants of EREG were coexpressed with FLAG-EGFR in HEK293 cells. EREG and EGFR were immunoprecipitated with anti-HA and anti-FLAG antibodies, respectively, and analyzed by Western blotting.

    Techniques Used: Immunoprecipitation, Western Blot, Immunofluorescence, Staining, Mutagenesis, Construct, Sequencing

    EGFR-Erk activation by epiregulin is sustained. (A) Representative time courses of EGFR phosphorylation at Y1086 in SACC-83 cells induced by EGF, EREG, AREG or TGF-α. An anti-EGFR antibody was used as a loading control. (B) Representative time courses of EGFR phosphorylation at Y1173 in CAL27 cells induced by EGF, EREG, AREG or TGF-α. An anti-EGFR antibody was used as a loading control. (C-D) Quantification of EGFR phosphorylation time courses, normalized by the signal at 5 min. The data are plotted on the same graph for multiple independent experiments quantitating phosphorylation at Y1068 and Y1173. (E-F) Representative time courses of Erk phosphorylation in SACC-83 and CAL27 cells induced by different EGFR ligands. (G-H) Quantification of Erk phosphorylation time courses, normalized by the signal at 5 min. The data are plotted on the same graph for multiple independent experiments quantifying Erk phosphorylation.
    Figure Legend Snippet: EGFR-Erk activation by epiregulin is sustained. (A) Representative time courses of EGFR phosphorylation at Y1086 in SACC-83 cells induced by EGF, EREG, AREG or TGF-α. An anti-EGFR antibody was used as a loading control. (B) Representative time courses of EGFR phosphorylation at Y1173 in CAL27 cells induced by EGF, EREG, AREG or TGF-α. An anti-EGFR antibody was used as a loading control. (C-D) Quantification of EGFR phosphorylation time courses, normalized by the signal at 5 min. The data are plotted on the same graph for multiple independent experiments quantitating phosphorylation at Y1068 and Y1173. (E-F) Representative time courses of Erk phosphorylation in SACC-83 and CAL27 cells induced by different EGFR ligands. (G-H) Quantification of Erk phosphorylation time courses, normalized by the signal at 5 min. The data are plotted on the same graph for multiple independent experiments quantifying Erk phosphorylation.

    Techniques Used: Activation Assay

    EREG-induced C-Myc expression depends on EGFR activity. (A) Western blot analysis of C-Myc, p-EGFR, and EGFR from tumor cell lines pretreated with various EGFR inhibitors for 1 h followed by stimulation with epiregulin for 2 h. (B) OncoPrint of EREG-EGFR-MYC pathway alterations in HNC. Genomic alterations of different members of this pathway are mutually exclusive. (C) Western blot analysis of C-Myc, p-EGFR, EGFR, p-AKT, AKT, p-ERK, ERK, p-STAT3, and STAT3 from CAL27 and HN6 cells pretreated with various inhibitors for 1 h followed by stimulation with epiregulin for 2 h.
    Figure Legend Snippet: EREG-induced C-Myc expression depends on EGFR activity. (A) Western blot analysis of C-Myc, p-EGFR, and EGFR from tumor cell lines pretreated with various EGFR inhibitors for 1 h followed by stimulation with epiregulin for 2 h. (B) OncoPrint of EREG-EGFR-MYC pathway alterations in HNC. Genomic alterations of different members of this pathway are mutually exclusive. (C) Western blot analysis of C-Myc, p-EGFR, EGFR, p-AKT, AKT, p-ERK, ERK, p-STAT3, and STAT3 from CAL27 and HN6 cells pretreated with various inhibitors for 1 h followed by stimulation with epiregulin for 2 h.

    Techniques Used: Expressing, Activity Assay, Western Blot

    EREG associates with EGFR and triggers EGFR signaling. (A) Immunoblot (IB) of HN13 and HN6 cells treated with epiregulin (50 ng/ml) at the indicated time points and probed with an anti-phosphotyrosine (p-Tyr) antibody. (B) IB of SACC and HNSCC cancer cells treated with epiregulin (50 ng/ml) for 5 min and probed with an anti-p-Tyr antibody. (C) Human phospho-RTK antibody array analysis of HN6 cells serum starved for 24 hr, followed by epiregulin (50 ng/mL) treatment for 5 min. (D) IB of HN13 (left) and HN6 (right) cells treated with epiregulin (50 ng/ml) at various time points. (E) Immunofluorescence staining for EGFR in HN6 cells treated with or without 50 ng/mL epiregulin.
    Figure Legend Snippet: EREG associates with EGFR and triggers EGFR signaling. (A) Immunoblot (IB) of HN13 and HN6 cells treated with epiregulin (50 ng/ml) at the indicated time points and probed with an anti-phosphotyrosine (p-Tyr) antibody. (B) IB of SACC and HNSCC cancer cells treated with epiregulin (50 ng/ml) for 5 min and probed with an anti-p-Tyr antibody. (C) Human phospho-RTK antibody array analysis of HN6 cells serum starved for 24 hr, followed by epiregulin (50 ng/mL) treatment for 5 min. (D) IB of HN13 (left) and HN6 (right) cells treated with epiregulin (50 ng/ml) at various time points. (E) Immunofluorescence staining for EGFR in HN6 cells treated with or without 50 ng/mL epiregulin.

    Techniques Used: Ab Array, Immunofluorescence, Staining

    EREG triggers EGFR downstream signaling in an EGFR kinase-dependent manner. (A) HN13 and HN6 cells pretreated with erlotinib and AG1478 followed by epiregulin treatment and IB with the indicated antibodies. (B) IB of HN6 and HN13 cells transfected with individual small interfering RNAs (siRNAs) against EGFR in the presence or absence of epiregulin (50 ng/ml). (C) IB of HN6 and HN12 cells treated with or without epiregulin (50 ng/ml). (D) Human phosphokinase antibody array analysis of HN6 cells treated with or without epiregulin (50 ng/ml) for 5 min. (E) IB of HN13 (left) and HN6 (right) cells treated with epiregulin (50 ng/ml) at different time points. (F) HN6 and HN13 cells pretreated with erlotinib and AG1478 followed by epiregulin treatment and immunoblotting (IB) with the indicated antibodies.
    Figure Legend Snippet: EREG triggers EGFR downstream signaling in an EGFR kinase-dependent manner. (A) HN13 and HN6 cells pretreated with erlotinib and AG1478 followed by epiregulin treatment and IB with the indicated antibodies. (B) IB of HN6 and HN13 cells transfected with individual small interfering RNAs (siRNAs) against EGFR in the presence or absence of epiregulin (50 ng/ml). (C) IB of HN6 and HN12 cells treated with or without epiregulin (50 ng/ml). (D) Human phosphokinase antibody array analysis of HN6 cells treated with or without epiregulin (50 ng/ml) for 5 min. (E) IB of HN13 (left) and HN6 (right) cells treated with epiregulin (50 ng/ml) at different time points. (F) HN6 and HN13 cells pretreated with erlotinib and AG1478 followed by epiregulin treatment and immunoblotting (IB) with the indicated antibodies.

    Techniques Used: Transfection, Ab Array

    The proposed model showing that EGFR domains I and III and the N57 residue of EREG are required for EREG-induced EGFR-Erk-C-Myc signaling activation, which in turn promotes oncogenesis and increases erlotinib sensitivity in HNSCC patients.
    Figure Legend Snippet: The proposed model showing that EGFR domains I and III and the N57 residue of EREG are required for EREG-induced EGFR-Erk-C-Myc signaling activation, which in turn promotes oncogenesis and increases erlotinib sensitivity in HNSCC patients.

    Techniques Used: Activation Assay

    7) Product Images from "Positive crosstalk between EGFR and the TF-PAR2 pathway mediates resistance to cisplatin and poor survival in cervical cancer"

    Article Title: Positive crosstalk between EGFR and the TF-PAR2 pathway mediates resistance to cisplatin and poor survival in cervical cancer

    Journal: Oncotarget

    doi: 10.18632/oncotarget.25748

    PAR2 transactivates the EGFR-ERK signaling pathway in CASKI cells ( A ) CASKI cells were starved for 16 h followed by stimulation with PAR1-AP (50 µM), PAR2-AP (50 µM) or FVIIa (20 nM) for 15 minutes. After cell lysis, equal amounts of protein were used for the determination of p-EGFR using the PathScan Phospho-EGF Receptor (panTyr) Sandwich ELISA Kit. Values represent mean + SD of three independent experiments; ns: not significant, ** P
    Figure Legend Snippet: PAR2 transactivates the EGFR-ERK signaling pathway in CASKI cells ( A ) CASKI cells were starved for 16 h followed by stimulation with PAR1-AP (50 µM), PAR2-AP (50 µM) or FVIIa (20 nM) for 15 minutes. After cell lysis, equal amounts of protein were used for the determination of p-EGFR using the PathScan Phospho-EGF Receptor (panTyr) Sandwich ELISA Kit. Values represent mean + SD of three independent experiments; ns: not significant, ** P

    Techniques Used: Lysis, Sandwich ELISA

    EGFR activation upregulates TF in cervical cancer cells ( A ) CASKI cells were starved for 16 h and were treated with cetuximab (100 µg/mL). One hour later, cells were stimulated with EGF (50 ng/mL). After 10 min, cells were lysed, and the levels of p-EGFR (Tyr 1068) and HSC70 (loading control) were determined by Western blotting (representative image from three experiments). ( B ) After starving for 16 h, CASKI cells were treated with cetuximab (100 µg/mL). One hour later, cells were incubated with EGF (50 ng/mL). After treatment for 1.5 h, total RNA was extracted, and mRNA was converted into cDNA. Gene expression assay for TF ( F3 gene) was performed by quantitative PCR. GAPDH was used as a housekeeping gene. The relative expression level of mRNA was calculated using the ΔΔCT method. Values represent mean + SD of four independent experiments; *** P
    Figure Legend Snippet: EGFR activation upregulates TF in cervical cancer cells ( A ) CASKI cells were starved for 16 h and were treated with cetuximab (100 µg/mL). One hour later, cells were stimulated with EGF (50 ng/mL). After 10 min, cells were lysed, and the levels of p-EGFR (Tyr 1068) and HSC70 (loading control) were determined by Western blotting (representative image from three experiments). ( B ) After starving for 16 h, CASKI cells were treated with cetuximab (100 µg/mL). One hour later, cells were incubated with EGF (50 ng/mL). After treatment for 1.5 h, total RNA was extracted, and mRNA was converted into cDNA. Gene expression assay for TF ( F3 gene) was performed by quantitative PCR. GAPDH was used as a housekeeping gene. The relative expression level of mRNA was calculated using the ΔΔCT method. Values represent mean + SD of four independent experiments; *** P

    Techniques Used: Activation Assay, Western Blot, Incubation, Expressing, Real-time Polymerase Chain Reaction

    8) Product Images from "HCRP-1 regulates EGFR–AKT–BIM-mediated anoikis resistance and serves as a prognostic marker in human colon cancer"

    Article Title: HCRP-1 regulates EGFR–AKT–BIM-mediated anoikis resistance and serves as a prognostic marker in human colon cancer

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-1217-2

    Hypothetic model of HCRP-1 depletion inhibits BIM expression via the EGFR–AKT signaling pathway
    Figure Legend Snippet: Hypothetic model of HCRP-1 depletion inhibits BIM expression via the EGFR–AKT signaling pathway

    Techniques Used: Expressing

    Suppression of EGFR blocks the loss of HCRP-1 induced by the AKT signaling pathway, upregulates BIM expression, and suppresses anoikis resistance. a , b HCT116 and SW620 cells were pretreated with HCRP-1 siRNA and the EGFR inhibitor, AG1478 (5 mM/L), and then suspended in six-well plates with low-attachment surface. The whole-cell lysates were analyzed for the protein levels of p-EGFR, EGFR, p-AKT, AKT, BIM, and β-actin by using western blot. c , d Cell anoikis was detected by flow cytometry. HCT116 and SW620 cells were pretreated with HCRP-1 siRNA and the EGFR inhibitor, AG1478 (5 mM/L), and then suspended in six-well plates with low- attachment surface. Cells that did not adhere were subjected to flow cytometry detection. e and f HCT116 and SW620 cells were co-transfected with HCRP-1 siRNA and/or EGFR siRNA. Levels of p-EGFR, EGFR, p-AKT, AKT, BIM, and β-actin in cells that did not adhere were determined by Western blot. g , h Cell anoikis was detected by flow cytometry in colon cancer cells pretreated with HCRP-1 siRNA and/or EGFR siRNA. Error bars indicate mean ± SD. Every experiment was repeated at least three times. * P
    Figure Legend Snippet: Suppression of EGFR blocks the loss of HCRP-1 induced by the AKT signaling pathway, upregulates BIM expression, and suppresses anoikis resistance. a , b HCT116 and SW620 cells were pretreated with HCRP-1 siRNA and the EGFR inhibitor, AG1478 (5 mM/L), and then suspended in six-well plates with low-attachment surface. The whole-cell lysates were analyzed for the protein levels of p-EGFR, EGFR, p-AKT, AKT, BIM, and β-actin by using western blot. c , d Cell anoikis was detected by flow cytometry. HCT116 and SW620 cells were pretreated with HCRP-1 siRNA and the EGFR inhibitor, AG1478 (5 mM/L), and then suspended in six-well plates with low- attachment surface. Cells that did not adhere were subjected to flow cytometry detection. e and f HCT116 and SW620 cells were co-transfected with HCRP-1 siRNA and/or EGFR siRNA. Levels of p-EGFR, EGFR, p-AKT, AKT, BIM, and β-actin in cells that did not adhere were determined by Western blot. g , h Cell anoikis was detected by flow cytometry in colon cancer cells pretreated with HCRP-1 siRNA and/or EGFR siRNA. Error bars indicate mean ± SD. Every experiment was repeated at least three times. * P

    Techniques Used: Expressing, Western Blot, Flow Cytometry, Cytometry, Transfection

    Inhibition of HCRP-1 activates the EGFR/AKT signaling pathway and suppresses BIM protein expression. a , b HCT116 and SW620 cells were transfected with si-HCRP-1 or control siRNA, and then cells were harvested and submitted to Western blot detection for the protein expression of BIM, FoxO3a, EGFR, AKT, ERK, Mcl-1, Bcl-2, and β-actin. Phosphorylated forms of EGFR, AKT, FoxO3a, and ERK were also detected by western blot with the corresponding antibodies. c–f Cells were treatedwith CHX for 0, 0.5 , 1, and 2 h after transfection with si-HCRP-1 or control siRNA for 48 h, lysates obtained from these cells were submitted to western blot detection for the protein expression of EGFR. Error bars indicate mean ± SD. Every experiment was repeated at least three times. * P
    Figure Legend Snippet: Inhibition of HCRP-1 activates the EGFR/AKT signaling pathway and suppresses BIM protein expression. a , b HCT116 and SW620 cells were transfected with si-HCRP-1 or control siRNA, and then cells were harvested and submitted to Western blot detection for the protein expression of BIM, FoxO3a, EGFR, AKT, ERK, Mcl-1, Bcl-2, and β-actin. Phosphorylated forms of EGFR, AKT, FoxO3a, and ERK were also detected by western blot with the corresponding antibodies. c–f Cells were treatedwith CHX for 0, 0.5 , 1, and 2 h after transfection with si-HCRP-1 or control siRNA for 48 h, lysates obtained from these cells were submitted to western blot detection for the protein expression of EGFR. Error bars indicate mean ± SD. Every experiment was repeated at least three times. * P

    Techniques Used: Inhibition, Expressing, Transfection, Western Blot

    9) Product Images from "Identification of epipolythiodioxopiperazines HDN-1 and chaetocin as novel inhibitor of heat shock protein 90"

    Article Title: Identification of epipolythiodioxopiperazines HDN-1 and chaetocin as novel inhibitor of heat shock protein 90

    Journal: Oncotarget

    doi:

    HDN-1 promotes EGF-induced EGFR downregulation (A) EGFR mutants in H1975 and HCC827 cells were resistant to be degraded upon EGF treatment. A549, H1975 or HCC827 cells were cultured overnight with serum free medium and incubated with 25 ng/ml EGF at indicated time. EGFR expression was determined by Western blotting assay. (B, C) HDN-1 promoted the reduction of EGFR expression and upregulation of EGFR Tyr1045 phosphorylation upon EGF stimulation in A549, H1975 and HCC827 cells. A549, H1975 or HCC827 cells were incubated with HDN-1 at indicated concentrations and time in the presence of 25 ng/ml EGF. EGFR and EGFR pY1045 were determined by Western blotting assay. (D) HDN-1 increased the level of EGFR ubiquitination in A549 cell. A549 cells were pretreated with 10 μM MG132 for 1 h, and then incubated with HDN-1 at indicated concentrations in the presence with 25 ng/ml EGF for 30 min. The levels of EGFR ubiquitination were detected by an antibody for ubiquitin.
    Figure Legend Snippet: HDN-1 promotes EGF-induced EGFR downregulation (A) EGFR mutants in H1975 and HCC827 cells were resistant to be degraded upon EGF treatment. A549, H1975 or HCC827 cells were cultured overnight with serum free medium and incubated with 25 ng/ml EGF at indicated time. EGFR expression was determined by Western blotting assay. (B, C) HDN-1 promoted the reduction of EGFR expression and upregulation of EGFR Tyr1045 phosphorylation upon EGF stimulation in A549, H1975 and HCC827 cells. A549, H1975 or HCC827 cells were incubated with HDN-1 at indicated concentrations and time in the presence of 25 ng/ml EGF. EGFR and EGFR pY1045 were determined by Western blotting assay. (D) HDN-1 increased the level of EGFR ubiquitination in A549 cell. A549 cells were pretreated with 10 μM MG132 for 1 h, and then incubated with HDN-1 at indicated concentrations in the presence with 25 ng/ml EGF for 30 min. The levels of EGFR ubiquitination were detected by an antibody for ubiquitin.

    Techniques Used: Cell Culture, Incubation, Expressing, Western Blot

    10) Product Images from "Combinational therapy of crizotinib and afatinib for malignant pleural mesothelioma"

    Article Title: Combinational therapy of crizotinib and afatinib for malignant pleural mesothelioma

    Journal: American Journal of Cancer Research

    doi:

    Simultaneous treatment with MET and EGFR inhibitors induced remarkably greater anticancer effect on MPM cell xenografts in vivo . A. The changes of tumor volume over time following the implantation. Data points represented the mean ± SD of tumor
    Figure Legend Snippet: Simultaneous treatment with MET and EGFR inhibitors induced remarkably greater anticancer effect on MPM cell xenografts in vivo . A. The changes of tumor volume over time following the implantation. Data points represented the mean ± SD of tumor

    Techniques Used: In Vivo

    Kaplan-Meier curve for overall survival of MPM patients with expression of MET and EGFR. The overall survival rates were estimated by the Kaplan-Meier method, and the differences between curves were tested using the log-rank test. A. A difference in overall
    Figure Legend Snippet: Kaplan-Meier curve for overall survival of MPM patients with expression of MET and EGFR. The overall survival rates were estimated by the Kaplan-Meier method, and the differences between curves were tested using the log-rank test. A. A difference in overall

    Techniques Used: Expressing

    Simultaneous treatment with MET and EGFR inhibitors induced remarkably inhibitory effect on MPM cell proliferation in vitro
    Figure Legend Snippet: Simultaneous treatment with MET and EGFR inhibitors induced remarkably inhibitory effect on MPM cell proliferation in vitro

    Techniques Used: In Vitro

    Simultaneous treatment with MET and EGFR inhibitors induced significantly anticancer effect on MPM cell proliferation in vitro . A. Five MPM cell lines express wild-type EGFR validated by DNA sequencing. B, C. Cells were treated with crizotinib or afatinib
    Figure Legend Snippet: Simultaneous treatment with MET and EGFR inhibitors induced significantly anticancer effect on MPM cell proliferation in vitro . A. Five MPM cell lines express wild-type EGFR validated by DNA sequencing. B, C. Cells were treated with crizotinib or afatinib

    Techniques Used: In Vitro, DNA Sequencing

    Downstream of MET and EGFR was active in MPM cells and specimens. A. Representative graphs showing typical examples of the four intensity grades of p-MET, p-EGFR, p-AKT and p-ERK staining in tumor samples of the cohort of 24 MPM patients. B. Statistics
    Figure Legend Snippet: Downstream of MET and EGFR was active in MPM cells and specimens. A. Representative graphs showing typical examples of the four intensity grades of p-MET, p-EGFR, p-AKT and p-ERK staining in tumor samples of the cohort of 24 MPM patients. B. Statistics

    Techniques Used: Staining

    Simutaneously suppress MET and EGFR can remarkly inhibit MPM cells proliferation. A and C. MET and EGFR protein levels were detected by Western Blotting after transfection of MET-siRNA and EGFR-siRNA. GAPDH was used as a loading control. The data represent
    Figure Legend Snippet: Simutaneously suppress MET and EGFR can remarkly inhibit MPM cells proliferation. A and C. MET and EGFR protein levels were detected by Western Blotting after transfection of MET-siRNA and EGFR-siRNA. GAPDH was used as a loading control. The data represent

    Techniques Used: Western Blot, Transfection

    Simultaneous treatment with MET and EGFR inhibitors induced remarkably inhibitory effect on MPM cell proliferation in vitro
    Figure Legend Snippet: Simultaneous treatment with MET and EGFR inhibitors induced remarkably inhibitory effect on MPM cell proliferation in vitro

    Techniques Used: In Vitro

    11) Product Images from "Cartilage-specific deletion of Mig-6 results in osteoarthritis-like disorder with excessive articular chondrocyte proliferation"

    Article Title: Cartilage-specific deletion of Mig-6 results in osteoarthritis-like disorder with excessive articular chondrocyte proliferation

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1400744111

    Detection of collagens type II, type X, PCNA, p-EGFR, and p-ERK in the Mig-6 flox/flox ;Col2a1 Cre knee joint. Strong staining of collagen II was detected within the osteophytes (labeled “nod”) in both 3-mo-old ( A ) and 15.3-mo-old ( B ) knee
    Figure Legend Snippet: Detection of collagens type II, type X, PCNA, p-EGFR, and p-ERK in the Mig-6 flox/flox ;Col2a1 Cre knee joint. Strong staining of collagen II was detected within the osteophytes (labeled “nod”) in both 3-mo-old ( A ) and 15.3-mo-old ( B ) knee

    Techniques Used: Staining, Labeling

    12) Product Images from "PGE2/EP4 receptor attenuated mucosal injury via β‐arrestin1/Src/EGFR‐mediated proliferation in portal hypertensive gastropathy) PGE2/EP4 receptor attenuated mucosal injury via β‐arrestin1/Src/EGFR‐mediated proliferation in portal hypertensive gastropathy"

    Article Title: PGE2/EP4 receptor attenuated mucosal injury via β‐arrestin1/Src/EGFR‐mediated proliferation in portal hypertensive gastropathy) PGE2/EP4 receptor attenuated mucosal injury via β‐arrestin1/Src/EGFR‐mediated proliferation in portal hypertensive gastropathy

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.13752

    Src and EGFR activations are suppressed in PHG. (A) Immunohistochemical staining for p‐EGFR and p‐Src from the representative uninvolved normal gastric mucosal tissue (from healthy volunteers) and gastropathic mucosal tissue from PHG patients is presented (brown, ×400, n = 6 per group). (B) Western blotting showed that p‐Src and p‐EGFR were repressed in PHG sections ( n = 6 per group). (C) and (D) The ratio of the normalized p‐Src/β‐actin and p‐EGFR/β‐actin was represented from western blotting ( n = 6 per group, Bonferroni's comparison post hoc test). (E) Immunofluorescence staining for p‐EGFR (red) with DAPI (blue) counterstaining for DNA in the indicated representative sections from PVL‐treated β‐arr1 ‐WT and β‐arr1 ‐KO mice, with or without PGE 2 treatment (3 mg·kg −1 ) (×400, n = 6 per group). (F) Immunofluorescence staining of p‐Src (red) was presented in the representative gastric mucosa of PVL‐treated mice; nuclei (blue) were counterstained with DAPI (×400, n = 6 per group).
    Figure Legend Snippet: Src and EGFR activations are suppressed in PHG. (A) Immunohistochemical staining for p‐EGFR and p‐Src from the representative uninvolved normal gastric mucosal tissue (from healthy volunteers) and gastropathic mucosal tissue from PHG patients is presented (brown, ×400, n = 6 per group). (B) Western blotting showed that p‐Src and p‐EGFR were repressed in PHG sections ( n = 6 per group). (C) and (D) The ratio of the normalized p‐Src/β‐actin and p‐EGFR/β‐actin was represented from western blotting ( n = 6 per group, Bonferroni's comparison post hoc test). (E) Immunofluorescence staining for p‐EGFR (red) with DAPI (blue) counterstaining for DNA in the indicated representative sections from PVL‐treated β‐arr1 ‐WT and β‐arr1 ‐KO mice, with or without PGE 2 treatment (3 mg·kg −1 ) (×400, n = 6 per group). (F) Immunofluorescence staining of p‐Src (red) was presented in the representative gastric mucosa of PVL‐treated mice; nuclei (blue) were counterstained with DAPI (×400, n = 6 per group).

    Techniques Used: Immunohistochemistry, Staining, Western Blot, Immunofluorescence, Mouse Assay

    PGE 2 /EP 4 receptor promotes Src and EGFR activation and complex formation via β‐arr1 in PHG. (A) Expression of the indicated proteins in the gastric mucosa of PVL‐treated β‐arr1 ‐WT and β‐arr1 ‐KO mice with or without PGE 2 treatment (3 mg·kg −1 ), as determined by western blotting. β‐actin was used as the loading control. n = 6 per group. (B) and (C) The ratio of densitometry units of the normalized p‐Src/β‐actin and p‐EGFR/β‐actin (EGFR) was represented from western blotting ( n = 6 per group, Bonferroni's comparison post hoc test). * P
    Figure Legend Snippet: PGE 2 /EP 4 receptor promotes Src and EGFR activation and complex formation via β‐arr1 in PHG. (A) Expression of the indicated proteins in the gastric mucosa of PVL‐treated β‐arr1 ‐WT and β‐arr1 ‐KO mice with or without PGE 2 treatment (3 mg·kg −1 ), as determined by western blotting. β‐actin was used as the loading control. n = 6 per group. (B) and (C) The ratio of densitometry units of the normalized p‐Src/β‐actin and p‐EGFR/β‐actin (EGFR) was represented from western blotting ( n = 6 per group, Bonferroni's comparison post hoc test). * P

    Techniques Used: Activation Assay, Expressing, Mouse Assay, Western Blot

    Inhibition of Src or EGFR blocks PGE 2 /EP 4 receptor‐mediated mucosal proliferation in PHG. (A) The Src inhibitor PP2 (80 nmol per mouse) significantly suppressed PVL‐induced p‐Src and p‐EGFR expression in mice, as determined by western blotting. n = 6 per group. (B) Western blotting revealed that the EGFR inhibitor AG1478 (80 nmol per mouse) suppressed PVL‐induced p‐EGFR expression in both PGE 2 ‐treated β‐arr1 ‐WT and β‐arr1 ‐KO mice under PVL. n = 6 per group. (C) and (D) PCNA‐stained images of the groups indicated (brown, ×200, n = 6 per group). (E) and (F) The proliferation index from PCNA staining was analysed as described previously. * P
    Figure Legend Snippet: Inhibition of Src or EGFR blocks PGE 2 /EP 4 receptor‐mediated mucosal proliferation in PHG. (A) The Src inhibitor PP2 (80 nmol per mouse) significantly suppressed PVL‐induced p‐Src and p‐EGFR expression in mice, as determined by western blotting. n = 6 per group. (B) Western blotting revealed that the EGFR inhibitor AG1478 (80 nmol per mouse) suppressed PVL‐induced p‐EGFR expression in both PGE 2 ‐treated β‐arr1 ‐WT and β‐arr1 ‐KO mice under PVL. n = 6 per group. (C) and (D) PCNA‐stained images of the groups indicated (brown, ×200, n = 6 per group). (E) and (F) The proliferation index from PCNA staining was analysed as described previously. * P

    Techniques Used: Inhibition, Expressing, Mouse Assay, Western Blot, Staining

    The PGE 2 /EP 4 receptor‐mediated β‐arr1/Src/EGFR complex contributes to mucosal proliferation via Akt activation in PHG. (A) PP2 treatment (80 nmol per mouse) blocked the upregulation of p‐Akt and PCNA in β‐arr1 ‐WT mice following PGE 2 administration (3 mg·kg −1 ) and promoted the expression of cleaved caspase‐3 in the representative mucosa of β‐arr1 ‐WT mice following PGE 2 administration. β‐actin was used as the loading control, n = 6 per group. (B) AG1478 treatment (80 nmol per mouse) down‐regulated the levels of p‐Akt and PCNA in β‐arr1 ‐WT mice following PGE 2 administration (3 mg·kg −1 ) and enhanced the expression of cleaved caspase‐3 in the mucosa of PVL‐treated β‐arr1 ‐WT mice with PGE 2 administration. β‐actin was used as the loading control, n = 6 per group. (C) and (D) The ratio of densitometry units of the normalized p‐Akt/β‐actin and PCNA/β‐actin of PP2‐treated sections from western blotting. * P
    Figure Legend Snippet: The PGE 2 /EP 4 receptor‐mediated β‐arr1/Src/EGFR complex contributes to mucosal proliferation via Akt activation in PHG. (A) PP2 treatment (80 nmol per mouse) blocked the upregulation of p‐Akt and PCNA in β‐arr1 ‐WT mice following PGE 2 administration (3 mg·kg −1 ) and promoted the expression of cleaved caspase‐3 in the representative mucosa of β‐arr1 ‐WT mice following PGE 2 administration. β‐actin was used as the loading control, n = 6 per group. (B) AG1478 treatment (80 nmol per mouse) down‐regulated the levels of p‐Akt and PCNA in β‐arr1 ‐WT mice following PGE 2 administration (3 mg·kg −1 ) and enhanced the expression of cleaved caspase‐3 in the mucosa of PVL‐treated β‐arr1 ‐WT mice with PGE 2 administration. β‐actin was used as the loading control, n = 6 per group. (C) and (D) The ratio of densitometry units of the normalized p‐Akt/β‐actin and PCNA/β‐actin of PP2‐treated sections from western blotting. * P

    Techniques Used: Activation Assay, Mouse Assay, Expressing, Western Blot

    Inhibition of Akt activation blocks β‐arr1/Src/EGFR‐mediated mucosal proliferation in PHG. (A) Treatment with the Akt inhibitor VIII (A6730, 50 mg·kg −1 ) down‐regulated p‐Akt and PCNA expression in β‐arr1 ‐WT mice following PGE 2 administration (3 mg·kg −1 ) but up‐regulated cleaved caspase‐3 expression in the mucosa of PVL‐treated β‐arr1 ‐WT mice administered PGE 2 . β‐actin was used as the loading control, n = 6 per group. (B) The Akt inhibitor VIII (50 mg·kg −1 ) did not affect the levels of p‐Src and p‐EGFR in PVL‐treated β‐arr1 ‐WT and β‐arr1 ‐KO mice following PGE 2 administration (3 mg·kg −1 ). β‐actin was used as the loading control, n = 6 per group. (C) and (D) The ratio of densitometry units of the normalized PCNA/β‐actin and cleaved caspase‐3/β‐actin of Akt inhibitor VIII‐treated sections from western blotting. * P
    Figure Legend Snippet: Inhibition of Akt activation blocks β‐arr1/Src/EGFR‐mediated mucosal proliferation in PHG. (A) Treatment with the Akt inhibitor VIII (A6730, 50 mg·kg −1 ) down‐regulated p‐Akt and PCNA expression in β‐arr1 ‐WT mice following PGE 2 administration (3 mg·kg −1 ) but up‐regulated cleaved caspase‐3 expression in the mucosa of PVL‐treated β‐arr1 ‐WT mice administered PGE 2 . β‐actin was used as the loading control, n = 6 per group. (B) The Akt inhibitor VIII (50 mg·kg −1 ) did not affect the levels of p‐Src and p‐EGFR in PVL‐treated β‐arr1 ‐WT and β‐arr1 ‐KO mice following PGE 2 administration (3 mg·kg −1 ). β‐actin was used as the loading control, n = 6 per group. (C) and (D) The ratio of densitometry units of the normalized PCNA/β‐actin and cleaved caspase‐3/β‐actin of Akt inhibitor VIII‐treated sections from western blotting. * P

    Techniques Used: Inhibition, Activation Assay, Expressing, Mouse Assay, Western Blot

    13) Product Images from "Tuberatolide B Suppresses Cancer Progression by Promoting ROS-Mediated Inhibition of STAT3 Signaling"

    Article Title: Tuberatolide B Suppresses Cancer Progression by Promoting ROS-Mediated Inhibition of STAT3 Signaling

    Journal: Marine Drugs

    doi: 10.3390/md15030055

    TTB selectively suppresses STAT3 signaling pathway: ( A ) MDA-MB-231, A549 and HCT116 cells were treated with TTB (100 μM) for 15 min and whole lysates were analyzed by western blot with anti-p-EGFR (Y992), -p-EGFR (Y1068), -p-EGFR (Y1173), -EGFR, -p-AKT, -AKT, -p-ERK, -ERK, -p-JNK, -JNK, -p-p38, -p38, -p-STAT3 and -STAT3. Actin was used for internal control. Quantitative analyses of p-STAT3 expression was performed using the Image J software; ( B ) MDA-MB-231 cells were transfected with the STAT3-dependent luciferase reporter and then treated with TTB (100 μM) for 6 h. Luciferase assay were done by using dual-luciferase reporter assays. All transfections included the RLTK-Luc for transfection efficiency. * p
    Figure Legend Snippet: TTB selectively suppresses STAT3 signaling pathway: ( A ) MDA-MB-231, A549 and HCT116 cells were treated with TTB (100 μM) for 15 min and whole lysates were analyzed by western blot with anti-p-EGFR (Y992), -p-EGFR (Y1068), -p-EGFR (Y1173), -EGFR, -p-AKT, -AKT, -p-ERK, -ERK, -p-JNK, -JNK, -p-p38, -p38, -p-STAT3 and -STAT3. Actin was used for internal control. Quantitative analyses of p-STAT3 expression was performed using the Image J software; ( B ) MDA-MB-231 cells were transfected with the STAT3-dependent luciferase reporter and then treated with TTB (100 μM) for 6 h. Luciferase assay were done by using dual-luciferase reporter assays. All transfections included the RLTK-Luc for transfection efficiency. * p

    Techniques Used: Multiple Displacement Amplification, Western Blot, Expressing, Software, Transfection, Luciferase

    14) Product Images from "The TWEAK Receptor Fn14 is a Src-inducible Protein and a Positive Regulator of Src-driven Cell Invasion"

    Article Title: The TWEAK Receptor Fn14 is a Src-inducible Protein and a Positive Regulator of Src-driven Cell Invasion

    Journal: Molecular cancer research : MCR

    doi: 10.1158/1541-7786.MCR-14-0411

    Effect of dasatinib treatment on Fn14 protein levels in EGFR-mutant HCC2279 and H1975 cells and EGF-stimulated, EGFR wild-type A549 cells
    Figure Legend Snippet: Effect of dasatinib treatment on Fn14 protein levels in EGFR-mutant HCC2279 and H1975 cells and EGF-stimulated, EGFR wild-type A549 cells

    Techniques Used: Mutagenesis

    Effect of erlotinib or signaling pathway inhibitor treatment on EGFR-driven Fn14 expression in HCC827 cells
    Figure Legend Snippet: Effect of erlotinib or signaling pathway inhibitor treatment on EGFR-driven Fn14 expression in HCC827 cells

    Techniques Used: Expressing

    15) Product Images from "Blockade of enhancer of zeste homolog 2 alleviates renal injury associated with hyperuricemia"

    Article Title: Blockade of enhancer of zeste homolog 2 alleviates renal injury associated with hyperuricemia

    Journal: American Journal of Physiology - Renal Physiology

    doi: 10.1152/ajprenal.00234.2018

    Small interfering RNA (siRNA)-mediated silencing of enhancer of zeste homolog 2 (EZH2) inhibits the activation of rat renal interstitial fibroblasts as well as the phosphorylation of Smad3, epidermal growth factor receptor (EGFR), and ERK1/2 in NRK-49F. Serum-starved NRK-49F cells were transferred with siRNA targeting EZH2 or scrambled siRNA and then incubated with uric acid (UA; 800 μM) for an additional 36 h. Cell lysates were subjected to immunoblot analysis with antibodies against α-smooth muscle actin (α-SMA), collagen I, EZH2, trimethylation of histone H3 at lysine27 (H3K27me3), p-Smad3, Smad3, p-EGFR, EGFR, p-ERK1/2, ERK1/2, or β-actin ( A , C , E , and G ). Expression levels of α-SMA, collagen I ( B ), and EZH2, H3K27me3 ( D ) were quantified by densitometry and normalized with β-actin. Expression of p-Smad3 was quantified by densitometry and normalized with Smad3 ( F ). Expression of p-EGFR was quantified by densitometry and normalized with EGFR ( H ). Expression of p-ERK1/2 was quantified by densitometry and normalized with ERK1/2 ( H ). Values are means ± SD of at least 3 independent experiments. Means with different letters (a–d) for each molecule are significantly different from one another ( P
    Figure Legend Snippet: Small interfering RNA (siRNA)-mediated silencing of enhancer of zeste homolog 2 (EZH2) inhibits the activation of rat renal interstitial fibroblasts as well as the phosphorylation of Smad3, epidermal growth factor receptor (EGFR), and ERK1/2 in NRK-49F. Serum-starved NRK-49F cells were transferred with siRNA targeting EZH2 or scrambled siRNA and then incubated with uric acid (UA; 800 μM) for an additional 36 h. Cell lysates were subjected to immunoblot analysis with antibodies against α-smooth muscle actin (α-SMA), collagen I, EZH2, trimethylation of histone H3 at lysine27 (H3K27me3), p-Smad3, Smad3, p-EGFR, EGFR, p-ERK1/2, ERK1/2, or β-actin ( A , C , E , and G ). Expression levels of α-SMA, collagen I ( B ), and EZH2, H3K27me3 ( D ) were quantified by densitometry and normalized with β-actin. Expression of p-Smad3 was quantified by densitometry and normalized with Smad3 ( F ). Expression of p-EGFR was quantified by densitometry and normalized with EGFR ( H ). Expression of p-ERK1/2 was quantified by densitometry and normalized with ERK1/2 ( H ). Values are means ± SD of at least 3 independent experiments. Means with different letters (a–d) for each molecule are significantly different from one another ( P

    Techniques Used: Small Interfering RNA, Activation Assay, Incubation, Expressing

    Inhibition of enhancer of zeste homolog 2 (EZH2) abrogates activation of epidermal growth factor receptor (EGFR)/ERK1/2 signaling in the kidney of hyperuricemia mice. A mouse model of hyperuricemia was established by intraperitoneal injection with uric acid (UA; 250 mg/kg) daily. The kidney tissue lysates were subjected to immunoblot analysis with specific antibodies against EGFR, p-EGFR, or β-actin ( A ). Expression level of p-EGFR was quantified by densitometry and normalized with EGFR ( B ). Expression level of EGFR was quantified by densitometry and normalized with β-actin ( C ). The kidney tissue lysates were subjected to immunoblot analysis with specific antibodies against ERK1/2, p-ERK1/2, or β-actin ( D ). Expression level of p-ERK1/2 was quantified by densitometry and normalized with ERK1/2 ( E ). Expression level of ERK1/2 was quantified by densitometry and normalized with β-actin ( F ). Data are represented as the means ± SD ( n = 6). Means with different letters (a, b) for each molecule are significantly different from one another ( P
    Figure Legend Snippet: Inhibition of enhancer of zeste homolog 2 (EZH2) abrogates activation of epidermal growth factor receptor (EGFR)/ERK1/2 signaling in the kidney of hyperuricemia mice. A mouse model of hyperuricemia was established by intraperitoneal injection with uric acid (UA; 250 mg/kg) daily. The kidney tissue lysates were subjected to immunoblot analysis with specific antibodies against EGFR, p-EGFR, or β-actin ( A ). Expression level of p-EGFR was quantified by densitometry and normalized with EGFR ( B ). Expression level of EGFR was quantified by densitometry and normalized with β-actin ( C ). The kidney tissue lysates were subjected to immunoblot analysis with specific antibodies against ERK1/2, p-ERK1/2, or β-actin ( D ). Expression level of p-ERK1/2 was quantified by densitometry and normalized with ERK1/2 ( E ). Expression level of ERK1/2 was quantified by densitometry and normalized with β-actin ( F ). Data are represented as the means ± SD ( n = 6). Means with different letters (a, b) for each molecule are significantly different from one another ( P

    Techniques Used: Inhibition, Activation Assay, Mouse Assay, Injection, Expressing

    Inhibition of enhancer of zeste homolog 2 (EZH2) suppresses phosphorylation of Smad3, epidermal growth factor receptor (EGFR), and ERK1/2 in NRK-49F. Serum-starved NRK-49F cells were pretreated with various concentration of 3-DZNeP (0, 1, 5, and 10 μM) for 1 h and then exposed to uric acid (UA; 800 μM) for an additional 36 h. Cell lysates were subjected to immunoblot analysis with antibodies against various molecules as indicated ( A , C , and E ). Expression of p-Smad3 was quantified by densitometry and normalized with Smad3 ( B ). Expression of p-EGFR was quantified by densitometry and normalized with EGFR ( D ). Expression of p-ERK1/2 was quantified by densitometry and normalized with ERK1/2 ( F ). Values are means ± SD of at least 3 independent experiments. Means with different letters (a–c) for each molecule are significantly different from one another ( P
    Figure Legend Snippet: Inhibition of enhancer of zeste homolog 2 (EZH2) suppresses phosphorylation of Smad3, epidermal growth factor receptor (EGFR), and ERK1/2 in NRK-49F. Serum-starved NRK-49F cells were pretreated with various concentration of 3-DZNeP (0, 1, 5, and 10 μM) for 1 h and then exposed to uric acid (UA; 800 μM) for an additional 36 h. Cell lysates were subjected to immunoblot analysis with antibodies against various molecules as indicated ( A , C , and E ). Expression of p-Smad3 was quantified by densitometry and normalized with Smad3 ( B ). Expression of p-EGFR was quantified by densitometry and normalized with EGFR ( D ). Expression of p-ERK1/2 was quantified by densitometry and normalized with ERK1/2 ( F ). Values are means ± SD of at least 3 independent experiments. Means with different letters (a–c) for each molecule are significantly different from one another ( P

    Techniques Used: Inhibition, Concentration Assay, Expressing

    16) Product Images from "METTL14 Inhibits Hepatocellular Carcinoma Metastasis Through Regulating EGFR/PI3K/AKT Signaling Pathway in an m6A-Dependent Manner"

    Article Title: METTL14 Inhibits Hepatocellular Carcinoma Metastasis Through Regulating EGFR/PI3K/AKT Signaling Pathway in an m6A-Dependent Manner

    Journal: Cancer Management and Research

    doi: 10.2147/CMAR.S286275

    Identifying EGFR as the target of METTL14 in HCC. ( A ) Analysis of RNA-sequencing and m6A-sequencing data of METTL14 knockdown in HepG2 cells deposited in GSE90642. ( B ) KEGG analysis of the potential targets of METTL14 in HCC. ( C) PPI analysis and the top 10 hub genes of the potential targets of METTL14. ( D ) METTL14 knockdown resulted in downregulated m6A levels but upregulated mRNA levels of EGFR in GSE90642. ( E ) The m6A peak in EGFR mRNA. ( F and G ) qPCR and Western blot analysis showed that METTL14 knockdown increased EGFR expression. ( H ) Western blot analysis showed that YTHDF2 knockdown increased EGFR expression. **p
    Figure Legend Snippet: Identifying EGFR as the target of METTL14 in HCC. ( A ) Analysis of RNA-sequencing and m6A-sequencing data of METTL14 knockdown in HepG2 cells deposited in GSE90642. ( B ) KEGG analysis of the potential targets of METTL14 in HCC. ( C) PPI analysis and the top 10 hub genes of the potential targets of METTL14. ( D ) METTL14 knockdown resulted in downregulated m6A levels but upregulated mRNA levels of EGFR in GSE90642. ( E ) The m6A peak in EGFR mRNA. ( F and G ) qPCR and Western blot analysis showed that METTL14 knockdown increased EGFR expression. ( H ) Western blot analysis showed that YTHDF2 knockdown increased EGFR expression. **p

    Techniques Used: RNA Sequencing Assay, Sequencing, Real-time Polymerase Chain Reaction, Western Blot, Expressing

    METTL14 inhibited the metastatic potential of HCC cells through regulating EGFR/PI3K/AKT signaling pathway. ( A ) Western blot analysis of the expression of EMT markers after siMETTL14 and siEGFR transfection. ( B ) Western blot analysis of the expression of p-EGFR, p-PI3K, and p-AKT after siMETTL14 and siEGFR transfection.
    Figure Legend Snippet: METTL14 inhibited the metastatic potential of HCC cells through regulating EGFR/PI3K/AKT signaling pathway. ( A ) Western blot analysis of the expression of EMT markers after siMETTL14 and siEGFR transfection. ( B ) Western blot analysis of the expression of p-EGFR, p-PI3K, and p-AKT after siMETTL14 and siEGFR transfection.

    Techniques Used: Western Blot, Expressing, Transfection

    EGFR knockdown effectively reversed the effect of METTL14 downregulation on HCC cells. ( A ) The transfection efficiency was tested by Western blot. ( B ) The wound healing assays showed that METTL14 knockdown promoted cell migration, whereas EGFR downregulation reversed such effect. ( C ) The Transwell Matrigel invasion assays showed that the improved invasive ability resulted by METTL14 knockdown was re-established after inhibiting EGFR expression in HCC cells. *p
    Figure Legend Snippet: EGFR knockdown effectively reversed the effect of METTL14 downregulation on HCC cells. ( A ) The transfection efficiency was tested by Western blot. ( B ) The wound healing assays showed that METTL14 knockdown promoted cell migration, whereas EGFR downregulation reversed such effect. ( C ) The Transwell Matrigel invasion assays showed that the improved invasive ability resulted by METTL14 knockdown was re-established after inhibiting EGFR expression in HCC cells. *p

    Techniques Used: Transfection, Western Blot, Migration, Expressing

    17) Product Images from "NFE2L2/NRF2 silencing-inducible miR-206 targets c-MET/EGFR and suppresses BCRP/ABCG2 in cancer cells"

    Article Title: NFE2L2/NRF2 silencing-inducible miR-206 targets c-MET/EGFR and suppresses BCRP/ABCG2 in cancer cells

    Journal: Oncotarget

    doi: 10.18632/oncotarget.22513

    The association of NRF2 with miR-206 is cancer cell type-dependent (A-B) c-MET and EGFR protein levels were determined in NRF2 -silenced HT29 (shNRF2-HT29; A) and NRF2 -silenced MDA-MB231 (shNRF2-MB231; B) cells. (C-D) MiR-206 levels were assessed by real-time PCR analysis in shNRF2-HT29 (C) and shNRF2-MB-231 cells (D). Values are means ± SD from three experiments. (E-F) After transfection of the scHT29 (E) and scMB-231 (F) with the miR-206 mimic (100 nM), protein levels for c-MET and EGFR were assessed by Western blotting. Similar blots were obtained from three independent experiments (A, B, E, and F).
    Figure Legend Snippet: The association of NRF2 with miR-206 is cancer cell type-dependent (A-B) c-MET and EGFR protein levels were determined in NRF2 -silenced HT29 (shNRF2-HT29; A) and NRF2 -silenced MDA-MB231 (shNRF2-MB231; B) cells. (C-D) MiR-206 levels were assessed by real-time PCR analysis in shNRF2-HT29 (C) and shNRF2-MB-231 cells (D). Values are means ± SD from three experiments. (E-F) After transfection of the scHT29 (E) and scMB-231 (F) with the miR-206 mimic (100 nM), protein levels for c-MET and EGFR were assessed by Western blotting. Similar blots were obtained from three independent experiments (A, B, E, and F).

    Techniques Used: Multiple Displacement Amplification, Real-time Polymerase Chain Reaction, Transfection, Western Blot

    MiR-206 mimic treatment represses c-MET and EGFR levels (A) The scSKOV3 cells were transfected with miR-206 mimic (100 nM) or negative control (NC), and c-MET, EGFR and GAPDH protein levels were determined by Western blot. Quantified results are means ± SD from three independent experiments. (B) MiR-206 levels in the sc and shNRF2-SKOV3 were confirmed using relative real-time PCR analysis. Values are means ± SD from three experiments. (C) Protein levels for AKR1C1 and NQO-1 were assessed in miR-206 expressing SKOV3 cells. (D) c-MET, EGFR, and GAPDH protein levels were determined by Western blotting after transfection of scA498 cells with miR-206 mimic or negative control (NC). (E) MiR-206 levels in the sc and shNRF2-A498 cells were confirmed using relative real-time PCR analysis. Values are means ± SD from three experiments. Similar blots were obtained from three independent experiments (B, C, and D).
    Figure Legend Snippet: MiR-206 mimic treatment represses c-MET and EGFR levels (A) The scSKOV3 cells were transfected with miR-206 mimic (100 nM) or negative control (NC), and c-MET, EGFR and GAPDH protein levels were determined by Western blot. Quantified results are means ± SD from three independent experiments. (B) MiR-206 levels in the sc and shNRF2-SKOV3 were confirmed using relative real-time PCR analysis. Values are means ± SD from three experiments. (C) Protein levels for AKR1C1 and NQO-1 were assessed in miR-206 expressing SKOV3 cells. (D) c-MET, EGFR, and GAPDH protein levels were determined by Western blotting after transfection of scA498 cells with miR-206 mimic or negative control (NC). (E) MiR-206 levels in the sc and shNRF2-A498 cells were confirmed using relative real-time PCR analysis. Values are means ± SD from three experiments. Similar blots were obtained from three independent experiments (B, C, and D).

    Techniques Used: Transfection, Negative Control, Western Blot, Real-time Polymerase Chain Reaction, Expressing

    The NRF2 -silencing-induced miR-206 directly mediates c-MET and EGFR reductions (A-B) SKOV3 (A) or A498 (B) cells were co-transfected with miR-206 and the c-MET -3′-UTR- or EGFR -3′UTR-luciferase reporter plasmid. After 18 h, 3-UTR-derived luciferase activity was measured. Values are means ± SD from 4 experiments. (C-D) After transfection of shNRF2-SKOV3 (C) or shNRF2-A498 (D) cells with the miR-206 inhibitor (100 nM) or negative control (NC), c-MET and EGFR protein levels were assessed by Western blotting. Data are means ± SD from three independent experiments. (E) The scSKOV3 cells were incubated with trichostatin (TSA, 0.3 μM) for 18 h, and miR-206 levels were measured using real-time PCR analysis. Values are means ± SD from three experiments. (F-G) Protein levels for c-MET and EGFR were determined in SKOV3 (I) and A498 (J) following the incubation with TSA (0.3 μM) for 24 h. (H) The transcript level of HDAC2 was assessed in sc and shNRF2-SKOV3 cells. (I) Cytoplasmic (Cyto) and nuclear (Nuc) levels of HDAC2 were determined Western blotting. (J) The scSKOV3 cells were transiently transfected with nonspecific siRNA (siCTL) or HDAC2 -specific siRNA (siHDAC2) and the protein levels for c-MET and EGFR were assessed by Western blotting. Similar blots were obtained from three independent experiments (D, F, G, I and J).
    Figure Legend Snippet: The NRF2 -silencing-induced miR-206 directly mediates c-MET and EGFR reductions (A-B) SKOV3 (A) or A498 (B) cells were co-transfected with miR-206 and the c-MET -3′-UTR- or EGFR -3′UTR-luciferase reporter plasmid. After 18 h, 3-UTR-derived luciferase activity was measured. Values are means ± SD from 4 experiments. (C-D) After transfection of shNRF2-SKOV3 (C) or shNRF2-A498 (D) cells with the miR-206 inhibitor (100 nM) or negative control (NC), c-MET and EGFR protein levels were assessed by Western blotting. Data are means ± SD from three independent experiments. (E) The scSKOV3 cells were incubated with trichostatin (TSA, 0.3 μM) for 18 h, and miR-206 levels were measured using real-time PCR analysis. Values are means ± SD from three experiments. (F-G) Protein levels for c-MET and EGFR were determined in SKOV3 (I) and A498 (J) following the incubation with TSA (0.3 μM) for 24 h. (H) The transcript level of HDAC2 was assessed in sc and shNRF2-SKOV3 cells. (I) Cytoplasmic (Cyto) and nuclear (Nuc) levels of HDAC2 were determined Western blotting. (J) The scSKOV3 cells were transiently transfected with nonspecific siRNA (siCTL) or HDAC2 -specific siRNA (siHDAC2) and the protein levels for c-MET and EGFR were assessed by Western blotting. Similar blots were obtained from three independent experiments (D, F, G, I and J).

    Techniques Used: Transfection, Luciferase, Plasmid Preparation, Derivative Assay, Activity Assay, Negative Control, Western Blot, Incubation, Real-time Polymerase Chain Reaction

    MiR-206 exhibits an antitumor effect in SKOV3 (A) SKOV3 cells were stably transduced with lentiviral particles containing miR-206- or negative control miRNA-expression plasmid. Levels of c-MET and EGFR protein were assessed by Western blotting in these two cell lines (SKOV3-CTRL and SKOV3-miR-206). (B) Proliferation rate of the SKOV3-CTRL and SKOV3-miR-206 cells was assessed using a TC10 Automated Cell Counter. Values are means ± SD from three experiments. (C) The SKOV3-CTRL and SKOV3-miR-206 were implanted in BALB/c-nu/nu mice and tumor growth was monitored weekly for 8 weeks. Tumor volume was calculated by the formula V = (a 2 x b)/2 (a is the width and b is the length in mm). Each group contained four to five animals. Values are means ± SE. Similar blots were obtained from three independent experiments (A).
    Figure Legend Snippet: MiR-206 exhibits an antitumor effect in SKOV3 (A) SKOV3 cells were stably transduced with lentiviral particles containing miR-206- or negative control miRNA-expression plasmid. Levels of c-MET and EGFR protein were assessed by Western blotting in these two cell lines (SKOV3-CTRL and SKOV3-miR-206). (B) Proliferation rate of the SKOV3-CTRL and SKOV3-miR-206 cells was assessed using a TC10 Automated Cell Counter. Values are means ± SD from three experiments. (C) The SKOV3-CTRL and SKOV3-miR-206 were implanted in BALB/c-nu/nu mice and tumor growth was monitored weekly for 8 weeks. Tumor volume was calculated by the formula V = (a 2 x b)/2 (a is the width and b is the length in mm). Each group contained four to five animals. Values are means ± SE. Similar blots were obtained from three independent experiments (A).

    Techniques Used: Stable Transfection, Transduction, Negative Control, Expressing, Plasmid Preparation, Western Blot, Mouse Assay

    The inhibition of c-MET or EGFR leads to BCRP reduction and doxorubicin sensitization (A) The scSKOV3 cells were incubated with PI3K inhibitor (LY294; 5 μM) and protein levels for BCRP were assessed by Western blotting. (B-C) After incubation of the scSKOV3 with pharmacological inhibitor of c-MET (SU112; 1 μM) or EGFR (LAPA; 2 μM, GEF; 5 μM), BCRP protein levels were determined. (D-E) The scSKOV3 cells were incubated with SU112 or LAPA, and cellular accumulation levels of Hoechst 33342 (H342; 2 μg/ml for 30 min; D) or doxorubicin (Dox; 2 μM for 6 h; E) were monitored. Cellular fluorescent intensities were quantified using a Cell Insight system. Values are means ± SD from 4 experiments. (F) The scSKOV3 cells were transiently transfected with c-MET -specific siRNA (siMET) or EGFR -specific siRNA (siEGFR), and the protein level for BCRP was assessed by Western blotting. (G) Transcript levels of BCRP were monitored using real-time PCR analysis after transfection with c-MET -specific siRNA (siMET) or EGFR -specific siRNA (siEGFR). Values are means ± SD from three experiments. Similar blots were obtained from three independent experiments (A, B, C and F).
    Figure Legend Snippet: The inhibition of c-MET or EGFR leads to BCRP reduction and doxorubicin sensitization (A) The scSKOV3 cells were incubated with PI3K inhibitor (LY294; 5 μM) and protein levels for BCRP were assessed by Western blotting. (B-C) After incubation of the scSKOV3 with pharmacological inhibitor of c-MET (SU112; 1 μM) or EGFR (LAPA; 2 μM, GEF; 5 μM), BCRP protein levels were determined. (D-E) The scSKOV3 cells were incubated with SU112 or LAPA, and cellular accumulation levels of Hoechst 33342 (H342; 2 μg/ml for 30 min; D) or doxorubicin (Dox; 2 μM for 6 h; E) were monitored. Cellular fluorescent intensities were quantified using a Cell Insight system. Values are means ± SD from 4 experiments. (F) The scSKOV3 cells were transiently transfected with c-MET -specific siRNA (siMET) or EGFR -specific siRNA (siEGFR), and the protein level for BCRP was assessed by Western blotting. (G) Transcript levels of BCRP were monitored using real-time PCR analysis after transfection with c-MET -specific siRNA (siMET) or EGFR -specific siRNA (siEGFR). Values are means ± SD from three experiments. Similar blots were obtained from three independent experiments (A, B, C and F).

    Techniques Used: Inhibition, Incubation, Western Blot, Transfection, Real-time Polymerase Chain Reaction

    NRF2 -silencing represses c-MET and EGFR levels in SKOV3 and A498 (A) The scSKOV3 and shNRF2-SKOV3 cell lines were established following stable transduction of cells with lentiviral particles containing either a nonspecific scRNA- or NRF2 shRNA-expressing plasmid. NRF2-silencing was verified by measuring protein levels for NRF2, AKR1C1, and NQO-1. (B) c-MET, EGFR, and GAPDH protein levels in sc and shNRF2-SKOV3 cells were examined by Western blotting. The quantified relative levels are means ± standard deviation (SD) from three independent experiments. (C) Transcript levels for c-MET and EGFR were determined by relative real-time PCR. (D) The sc and shNRF2-SKOV3 cells were incubated with serum-free media (SFM) for 24 h and protein levels for c-MET, p-c-MET (Tyr1234/1235), EGFR, p-EGFR (Tyr1068), p-AKT (Ser473), and p-ERK1/2 (Thr202/Tyr204) were determined by Western blotting. (E) HGF (10 ng/ml) was incubated for 24 h in scSKOV3 and shNRF2-SKOV3 cells, and levels for p-c-MET, p-AKT, and p-ERK1/2 were determined. (F) Lentiviral particles with pLKO-NRF2 plasmid were introduced into shNRF2-SKOV3 cells for forced expression of NRF2. Levels for NRF2, c-MET and EGFR were assessed in the control (pLKO) and NRF2 rescue group (pNRF2). (G) c-MET, EGFR and GAPDH protein levels in sc and shNRF2-A498 cells were examined. Values are means ± SD from three independent experiments. (H) Protein levels for c-MET, p-c-MET, EGFR, p-EGFR, p-AKT, and p-ERK1/2 were monitored under serum-free media (SFM) condition. Similar blots were obtained from three independent experiments (A, D, E, F, and H).
    Figure Legend Snippet: NRF2 -silencing represses c-MET and EGFR levels in SKOV3 and A498 (A) The scSKOV3 and shNRF2-SKOV3 cell lines were established following stable transduction of cells with lentiviral particles containing either a nonspecific scRNA- or NRF2 shRNA-expressing plasmid. NRF2-silencing was verified by measuring protein levels for NRF2, AKR1C1, and NQO-1. (B) c-MET, EGFR, and GAPDH protein levels in sc and shNRF2-SKOV3 cells were examined by Western blotting. The quantified relative levels are means ± standard deviation (SD) from three independent experiments. (C) Transcript levels for c-MET and EGFR were determined by relative real-time PCR. (D) The sc and shNRF2-SKOV3 cells were incubated with serum-free media (SFM) for 24 h and protein levels for c-MET, p-c-MET (Tyr1234/1235), EGFR, p-EGFR (Tyr1068), p-AKT (Ser473), and p-ERK1/2 (Thr202/Tyr204) were determined by Western blotting. (E) HGF (10 ng/ml) was incubated for 24 h in scSKOV3 and shNRF2-SKOV3 cells, and levels for p-c-MET, p-AKT, and p-ERK1/2 were determined. (F) Lentiviral particles with pLKO-NRF2 plasmid were introduced into shNRF2-SKOV3 cells for forced expression of NRF2. Levels for NRF2, c-MET and EGFR were assessed in the control (pLKO) and NRF2 rescue group (pNRF2). (G) c-MET, EGFR and GAPDH protein levels in sc and shNRF2-A498 cells were examined. Values are means ± SD from three independent experiments. (H) Protein levels for c-MET, p-c-MET, EGFR, p-EGFR, p-AKT, and p-ERK1/2 were monitored under serum-free media (SFM) condition. Similar blots were obtained from three independent experiments (A, D, E, F, and H).

    Techniques Used: Transduction, shRNA, Expressing, Plasmid Preparation, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction, Incubation

    18) Product Images from "Uropathogenic Escherichia coli invades bladder epithelial cells by activating kinase networks in host cells"

    Article Title: Uropathogenic Escherichia coli invades bladder epithelial cells by activating kinase networks in host cells

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.RA118.003499

    Role of EGFR, Akt, and mTORCs in UTI89 invasion of mouse bladder. A and B , strips of mouse transitional epithelial tissue were individually placed in wells (12-well plates) and treated or not with the indicated inhibitor for 1 h at 37 °C. UPEC (5 × 10 7 /ml) were added for additional 2 h, and tissue strips were washed three times with PBS. The tissue strips were lysed in PBS containing 0.1% (v/v) Triton X-100. For one part ( A ), cell lysates were analyzed for EGFR and Akt phosphorylation by Western blotting using the indicated antibodies. GAPDH was used as a protein loading control. For the second part ( B ), dilutions of lysates were plated on LB agar plates to allow colony growth. Colonies were counted, and data are presented as the -fold change of control values. Statistical analysis was performed by Student's t test compared with the nontreated ( NT ) control. *, p
    Figure Legend Snippet: Role of EGFR, Akt, and mTORCs in UTI89 invasion of mouse bladder. A and B , strips of mouse transitional epithelial tissue were individually placed in wells (12-well plates) and treated or not with the indicated inhibitor for 1 h at 37 °C. UPEC (5 × 10 7 /ml) were added for additional 2 h, and tissue strips were washed three times with PBS. The tissue strips were lysed in PBS containing 0.1% (v/v) Triton X-100. For one part ( A ), cell lysates were analyzed for EGFR and Akt phosphorylation by Western blotting using the indicated antibodies. GAPDH was used as a protein loading control. For the second part ( B ), dilutions of lysates were plated on LB agar plates to allow colony growth. Colonies were counted, and data are presented as the -fold change of control values. Statistical analysis was performed by Student's t test compared with the nontreated ( NT ) control. *, p

    Techniques Used: Western Blot

    19) Product Images from "microRNA-148a is a prognostic oncomiR that targets MIG6 and BIM to regulate EGFR and apoptosis in glioblastoma"

    Article Title: microRNA-148a is a prognostic oncomiR that targets MIG6 and BIM to regulate EGFR and apoptosis in glioblastoma

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-13-1449

    miR-148a directly targets and inhibits MIG6 and BIM and indirectly enhances EGFR expression and activation
    Figure Legend Snippet: miR-148a directly targets and inhibits MIG6 and BIM and indirectly enhances EGFR expression and activation

    Techniques Used: Expressing, Activation Assay

    20) Product Images from "EGFR targeting enhances the efficiency of chemotherapy through inhibiting IRE1α-XBP1s pathway in colorectal cancer cells"

    Article Title: EGFR targeting enhances the efficiency of chemotherapy through inhibiting IRE1α-XBP1s pathway in colorectal cancer cells

    Journal: Journal of Cancer

    doi: 10.7150/jca.44234

    EGFR activation is associated with IRE1α -XBP1s Signaling. A. Phosphorylation of EGFR and IRE1α were examined in SW480 cells 30min after stimulation of EGF (50ng/ml). B. Expression of spliced XBP1 mRNA in SW480 cells30 min after stimulation of EGF (50ng/ml). Values are represented as the mean ± SD (n = 3) for each treatment (*P
    Figure Legend Snippet: EGFR activation is associated with IRE1α -XBP1s Signaling. A. Phosphorylation of EGFR and IRE1α were examined in SW480 cells 30min after stimulation of EGF (50ng/ml). B. Expression of spliced XBP1 mRNA in SW480 cells30 min after stimulation of EGF (50ng/ml). Values are represented as the mean ± SD (n = 3) for each treatment (*P

    Techniques Used: Activation Assay, Expressing

    IRE1α-XBP1s pathway and EGFR expression in colorectal cancer. A. Immunoblotting of EGFR, IRE1α and XBP1s in paired tumor and normal tissues of colorectal cancer patients. GAPDH was used as a loading control. B. Expression of spliced XBP1 mRNA in 6 paired patient samples via q-PCR (n=6). Values are represented as the mean ± SD (n = 3) for each treatment (*P
    Figure Legend Snippet: IRE1α-XBP1s pathway and EGFR expression in colorectal cancer. A. Immunoblotting of EGFR, IRE1α and XBP1s in paired tumor and normal tissues of colorectal cancer patients. GAPDH was used as a loading control. B. Expression of spliced XBP1 mRNA in 6 paired patient samples via q-PCR (n=6). Values are represented as the mean ± SD (n = 3) for each treatment (*P

    Techniques Used: Expressing, Polymerase Chain Reaction

    EGFR signaling activates IRE1α through the kinase activity of ERK. A. The molecules of EGFR pathway were detected in SW480 cells by immunoblotting. B. The molecules of EGFR pathway of HCT116 EGFRKO cell line and HCT116 cells were examined by immunoblotting. C. EGFR downstream pathway were detected by immunoblotting in HCT116 treated with cetuximab (12.5μg/ml) for 24hrs. D. Proteins of IRE1α-XBP1s pathway in HCT116 cells with the treatment of MEK inhibitor PD0325901 (1nM) for 24hrs were assessed by immunoblotting. E. p-IRE1α (S724) and XBP1s were assessed via immunoblotting after the treatment of MEK inhibitor PD0325901 (1nM) or ERK inhibitor SCH8477 (5μM) for 24hrs. F. Expression of spliced XBP1 mRNA was assessed in SW480 cells after treatment with MEK inhibitor PD0325901 (1nM) or ERK inhibitor SCH8477 (5μM). MKC8866 was used as positive control. Values are represented as the mean ± SD (n = 3) for each treatment (****P
    Figure Legend Snippet: EGFR signaling activates IRE1α through the kinase activity of ERK. A. The molecules of EGFR pathway were detected in SW480 cells by immunoblotting. B. The molecules of EGFR pathway of HCT116 EGFRKO cell line and HCT116 cells were examined by immunoblotting. C. EGFR downstream pathway were detected by immunoblotting in HCT116 treated with cetuximab (12.5μg/ml) for 24hrs. D. Proteins of IRE1α-XBP1s pathway in HCT116 cells with the treatment of MEK inhibitor PD0325901 (1nM) for 24hrs were assessed by immunoblotting. E. p-IRE1α (S724) and XBP1s were assessed via immunoblotting after the treatment of MEK inhibitor PD0325901 (1nM) or ERK inhibitor SCH8477 (5μM) for 24hrs. F. Expression of spliced XBP1 mRNA was assessed in SW480 cells after treatment with MEK inhibitor PD0325901 (1nM) or ERK inhibitor SCH8477 (5μM). MKC8866 was used as positive control. Values are represented as the mean ± SD (n = 3) for each treatment (****P

    Techniques Used: Activity Assay, Expressing, Positive Control

    21) Product Images from "Inhibition of fucosylation by 2-fluorofucose suppresses human liver cancer HepG2 cell proliferation and migration as well as tumor formation"

    Article Title: Inhibition of fucosylation by 2-fluorofucose suppresses human liver cancer HepG2 cell proliferation and migration as well as tumor formation

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-11911-9

    Treatment with 2FF suppressed core fucosylation on EGFR and integrin β1, as well as down stream signaling. HepG2 cells were cultured with and without (control) 2FF for 3 days, and harvested for lectin pull down assay or immunoblotting as described in “Methods”. ( A ) The cell lysates were pooled and incubated with 10 μl of indicated bead-bound PhoSL, which specifically recognizes core fucose. The lectin precipitated samples and equal amounts of cell lysates were then subjected to SDS-PAGE, and probed with anti-EGFR (upper panel) and anti-integrin β1 (lower panel) antibodies. GAPDH were used as a loading control. ( B ) Cell lysates were immunoblotted with anti-phospho-EGFR, anti-EGFR, anti-phospho-AKT, anti-AKT, anti-phospho-ERK1/2, anti-ERK1/2, anti-phospho-FAK, and anti-FAK antibodies, respectively. GAPDH was used as a loading control.
    Figure Legend Snippet: Treatment with 2FF suppressed core fucosylation on EGFR and integrin β1, as well as down stream signaling. HepG2 cells were cultured with and without (control) 2FF for 3 days, and harvested for lectin pull down assay or immunoblotting as described in “Methods”. ( A ) The cell lysates were pooled and incubated with 10 μl of indicated bead-bound PhoSL, which specifically recognizes core fucose. The lectin precipitated samples and equal amounts of cell lysates were then subjected to SDS-PAGE, and probed with anti-EGFR (upper panel) and anti-integrin β1 (lower panel) antibodies. GAPDH were used as a loading control. ( B ) Cell lysates were immunoblotted with anti-phospho-EGFR, anti-EGFR, anti-phospho-AKT, anti-AKT, anti-phospho-ERK1/2, anti-ERK1/2, anti-phospho-FAK, and anti-FAK antibodies, respectively. GAPDH was used as a loading control.

    Techniques Used: Cell Culture, Pull Down Assay, Incubation, SDS Page

    22) Product Images from "Cetuximab and Cisplatin Show Different Combination Effect in Nasopharyngeal Carcinoma Cells Lines via Inactivation of EGFR/AKT Signaling Pathway"

    Article Title: Cetuximab and Cisplatin Show Different Combination Effect in Nasopharyngeal Carcinoma Cells Lines via Inactivation of EGFR/AKT Signaling Pathway

    Journal: Biochemistry Research International

    doi: 10.1155/2016/7016907

    Effects of combination treatment with cetuximab and cisplatin on the expressions of proapoptosis proteins and EGFR/AKT signaling pathway proteins. Cells were treated with cetuximab, cisplatin, or the combination treatment of cetuximab and cisplatin for 48 h. The expressions of EGFR, p-EGFR, AKT, p-AKT, Bax, caspase-3, and cleaved caspase-3 in both cells were detected by Western blotting analysis. β -actin was used as a loading control.
    Figure Legend Snippet: Effects of combination treatment with cetuximab and cisplatin on the expressions of proapoptosis proteins and EGFR/AKT signaling pathway proteins. Cells were treated with cetuximab, cisplatin, or the combination treatment of cetuximab and cisplatin for 48 h. The expressions of EGFR, p-EGFR, AKT, p-AKT, Bax, caspase-3, and cleaved caspase-3 in both cells were detected by Western blotting analysis. β -actin was used as a loading control.

    Techniques Used: Western Blot

    23) Product Images from "Transient IGF-1R inhibition combined with osimertinib eradicates AXL-low expressing EGFR mutated lung cancer"

    Article Title: Transient IGF-1R inhibition combined with osimertinib eradicates AXL-low expressing EGFR mutated lung cancer

    Journal: Nature Communications

    doi: 10.1038/s41467-020-18442-4

    IGF-1R inhibitor inhibited the viability of AXL-low tumor cells exposed to osimertinib. AXL-low expressing ( a ) and AXL-high expressing ( b ) EGFR -mutated NSCLC cell lines were treated with various concentrations of osimertinib for 72 h in the presence or absence of linsitinib (1 μmol/L), and cell viability was determined. The percentage of growth is shown relative to untreated controls. Data are presented as mean ± s.d. Each sample was assayed in triplicate, with each experiment repeated at least three times independently. c HCC4006 and HCC827 cells were treated with osimertinib (30 and 300 nmol/L, respectively), dacomitinib (30 and 300 nmol/L, respectively), and/or linsitinib (1 μmol/L). After 72 h, the cells were lysed, and the indicated proteins were detected by western blotting. Data shown are representative of three independent experiments. Bars indicate s.d.
    Figure Legend Snippet: IGF-1R inhibitor inhibited the viability of AXL-low tumor cells exposed to osimertinib. AXL-low expressing ( a ) and AXL-high expressing ( b ) EGFR -mutated NSCLC cell lines were treated with various concentrations of osimertinib for 72 h in the presence or absence of linsitinib (1 μmol/L), and cell viability was determined. The percentage of growth is shown relative to untreated controls. Data are presented as mean ± s.d. Each sample was assayed in triplicate, with each experiment repeated at least three times independently. c HCC4006 and HCC827 cells were treated with osimertinib (30 and 300 nmol/L, respectively), dacomitinib (30 and 300 nmol/L, respectively), and/or linsitinib (1 μmol/L). After 72 h, the cells were lysed, and the indicated proteins were detected by western blotting. Data shown are representative of three independent experiments. Bars indicate s.d.

    Techniques Used: Expressing, Western Blot

    AXL-low tumor cells show high level of pIGF-1R after osimertinib exposure. a EGFR -mutated NSCLC cells (seeded at 2 × 10 3 per well of a 96-well plate) were incubated with osimertinib at the indicated concentrations for 72 h. Cell viability was assessed using the Cell Counting Kit. Bars indicate s.d. of triplicate cultures. Data are presented as mean ± s.d. b Lysates of EGFR -mutated NSCLC cell lines were analyzed by western blotting. Data shown are representative of three independent experiments. c Correlation between the expression levels of the cytoplasmic AXL protein determined immunohistochemically, and response to treatment with osimertinib in EGFR -mutated NSCLC specimens from 29 patients. d Change in tumor size from baseline following osimertinib treatment in EGFR -mutated NSCLC patients with AXL-low ( n = 6) and AXL-high ( n = 23) expression. The data are expressed as mean and s.d. p value is provided (two-sided Student’s t -test). e Human tyrosine kinase phosphorylation array analysis in EGFR -mutated NSCLC cell lines in the presence or absence of osimertinib (30 nmol/L for HCC4006 and H3255 cells; 1 μmol/L for PC-9 cells) for 72 h. The circles indicate IGF-1R. f EGFR- mutated NSCLC cell lines were treated with osimertinib (30 nmol/L for HCC4006 and H3255 cells, 300 nmol/L for HCC827 cells) for indicated times, and lysates were analyzed by western blotting. Data shown are representative of three independent experiments.
    Figure Legend Snippet: AXL-low tumor cells show high level of pIGF-1R after osimertinib exposure. a EGFR -mutated NSCLC cells (seeded at 2 × 10 3 per well of a 96-well plate) were incubated with osimertinib at the indicated concentrations for 72 h. Cell viability was assessed using the Cell Counting Kit. Bars indicate s.d. of triplicate cultures. Data are presented as mean ± s.d. b Lysates of EGFR -mutated NSCLC cell lines were analyzed by western blotting. Data shown are representative of three independent experiments. c Correlation between the expression levels of the cytoplasmic AXL protein determined immunohistochemically, and response to treatment with osimertinib in EGFR -mutated NSCLC specimens from 29 patients. d Change in tumor size from baseline following osimertinib treatment in EGFR -mutated NSCLC patients with AXL-low ( n = 6) and AXL-high ( n = 23) expression. The data are expressed as mean and s.d. p value is provided (two-sided Student’s t -test). e Human tyrosine kinase phosphorylation array analysis in EGFR -mutated NSCLC cell lines in the presence or absence of osimertinib (30 nmol/L for HCC4006 and H3255 cells; 1 μmol/L for PC-9 cells) for 72 h. The circles indicate IGF-1R. f EGFR- mutated NSCLC cell lines were treated with osimertinib (30 nmol/L for HCC4006 and H3255 cells, 300 nmol/L for HCC827 cells) for indicated times, and lysates were analyzed by western blotting. Data shown are representative of three independent experiments.

    Techniques Used: Incubation, Cell Counting, Western Blot, Expressing

    pIGF-1R supports the survival of AXL-low tumor cells after osimertinib exposure. AXL-low expressing ( a ) and AXL-high expressing ( b ) EGFR -mutated NSCLC cell lines were treated with nonspecific control (si-Scr) or siRNA specific to IGF-1R (si-IGF-1R) for 72 h in the presence or absence of osimertinib (AXL-low of HCC4006 and HCC827:30 nmol/L and 300 nmol/L, respectively; AXL-high of PC-9 and HCC4011:1 μmol/L), and cell viability was determined. The percentage of growth is shown relative to untreated controls. Data are presented as mean ± s.d. Each sample was assayed in triplicate, with each experiment repeated at least three times independently. p values are provided (one-way ANOVA). n.s.: not significant. c si-Scr or si-IGF-1R was introduced into HCC4006 and HCC827 cells. After 24 h, the cells were incubated with or without osimertinib (30 nmol/L and 300 nmol/L, respectively) for 72 h and lysed, and the indicated proteins were detected by western blotting. d IGF-1R knockdown clones of HCC827 cells by CRISPR-CAS9 (KO-1-6, KO1-21, and KO2-14) were lysed and the proteins were detected by western blotting. e HCC827 and its IGF-1R knockdown clones were incubated with various concentrations of osimertinib, and cell viability was determined using the MTT assay. Data are presented as mean ± s.d. f HCC827 and KO1-6 clones were incubated with osimertinib (300 nmol/L) for 2 h, lysed, and the indicated proteins and their phosphorylation were detected by western blotting. Data shown are representative of three independent experiments.
    Figure Legend Snippet: pIGF-1R supports the survival of AXL-low tumor cells after osimertinib exposure. AXL-low expressing ( a ) and AXL-high expressing ( b ) EGFR -mutated NSCLC cell lines were treated with nonspecific control (si-Scr) or siRNA specific to IGF-1R (si-IGF-1R) for 72 h in the presence or absence of osimertinib (AXL-low of HCC4006 and HCC827:30 nmol/L and 300 nmol/L, respectively; AXL-high of PC-9 and HCC4011:1 μmol/L), and cell viability was determined. The percentage of growth is shown relative to untreated controls. Data are presented as mean ± s.d. Each sample was assayed in triplicate, with each experiment repeated at least three times independently. p values are provided (one-way ANOVA). n.s.: not significant. c si-Scr or si-IGF-1R was introduced into HCC4006 and HCC827 cells. After 24 h, the cells were incubated with or without osimertinib (30 nmol/L and 300 nmol/L, respectively) for 72 h and lysed, and the indicated proteins were detected by western blotting. d IGF-1R knockdown clones of HCC827 cells by CRISPR-CAS9 (KO-1-6, KO1-21, and KO2-14) were lysed and the proteins were detected by western blotting. e HCC827 and its IGF-1R knockdown clones were incubated with various concentrations of osimertinib, and cell viability was determined using the MTT assay. Data are presented as mean ± s.d. f HCC827 and KO1-6 clones were incubated with osimertinib (300 nmol/L) for 2 h, lysed, and the indicated proteins and their phosphorylation were detected by western blotting. Data shown are representative of three independent experiments.

    Techniques Used: Expressing, Incubation, Western Blot, Clone Assay, CRISPR, MTT Assay

    IGF-1R associated with EGFR and adaptor proteins, Gab1 and Shc. HCC4006 ( a ) and HCC827 ( b ) cells treated with osimertinib (30 nmol/L and 300 nmol/L, respectively) for 72 h were lysed, and the indicated proteins were detected by western blotting, with immunoprecipitation of the indicated proteins. c Schema presenting the mechanism by which AXL-low- expressing EGFR -mutated NSCLC cells adapt to osimertinib exposure. Data shown are representative of three independent experiments. IP immunoprecipitation.
    Figure Legend Snippet: IGF-1R associated with EGFR and adaptor proteins, Gab1 and Shc. HCC4006 ( a ) and HCC827 ( b ) cells treated with osimertinib (30 nmol/L and 300 nmol/L, respectively) for 72 h were lysed, and the indicated proteins were detected by western blotting, with immunoprecipitation of the indicated proteins. c Schema presenting the mechanism by which AXL-low- expressing EGFR -mutated NSCLC cells adapt to osimertinib exposure. Data shown are representative of three independent experiments. IP immunoprecipitation.

    Techniques Used: Western Blot, Immunoprecipitation, Expressing

    24) Product Images from "Discovery of WS-157 as a highly potent, selective and orally active EGFR inhibitor"

    Article Title: Discovery of WS-157 as a highly potent, selective and orally active EGFR inhibitor

    Journal: Acta Pharmaceutica Sinica. B

    doi: 10.1016/j.apsb.2019.06.010

    In vivo antitumor effects of compound WS-157 . Effect of WS-157 (10, 40 mg/kg/d) or gefitinib (10 mg/kg/d) on growth of A431 human tumor xenografts for 28 days. (A) Tumor volume during the administration period. (B) Tumor sizes. (C) Tumor weight. (D) Body weight of mice. (E) and (F) The tumor tissue protein was extracted and p-EGFR and p-AKT was detected by Western blot. The data shown represent the mean ± SD. ∗∗ P
    Figure Legend Snippet: In vivo antitumor effects of compound WS-157 . Effect of WS-157 (10, 40 mg/kg/d) or gefitinib (10 mg/kg/d) on growth of A431 human tumor xenografts for 28 days. (A) Tumor volume during the administration period. (B) Tumor sizes. (C) Tumor weight. (D) Body weight of mice. (E) and (F) The tumor tissue protein was extracted and p-EGFR and p-AKT was detected by Western blot. The data shown represent the mean ± SD. ∗∗ P

    Techniques Used: In Vivo, Mouse Assay, Western Blot

    25) Product Images from "Antitumor activity of the novel HDAC inhibitor CUDC-101 combined with gemcitabine in pancreatic cancer"

    Article Title: Antitumor activity of the novel HDAC inhibitor CUDC-101 combined with gemcitabine in pancreatic cancer

    Journal: American Journal of Cancer Research

    doi:

    Co-treatment with CUDC-101 and gemcitabine synergistically induced apoptosis signaling. A. Annexin V-FITC/PI stained of of PC cells after treatment with CUDC-101 and gemcitabine alone or combination for 48 h. Apoptotic cells were analyzed by flow cytometry. B. Western blot analysis showed CUDC-101 increased inhibition of gemcitabine on HDAC1, HDAC3, HDAC4 and p-EGFR proteins. Cyclin B1/CDK1 protein expression in CUDC-101 group was decreased. Acetylased histone H3 protein expression in CUDC-101 group and co-treatment group. C. Protein levels of p-p53, cl-caspase3 and the ratio of bax/bcl-2 were markedly increased in co-treatment group. Statistical significance was determined by a standard test (*P
    Figure Legend Snippet: Co-treatment with CUDC-101 and gemcitabine synergistically induced apoptosis signaling. A. Annexin V-FITC/PI stained of of PC cells after treatment with CUDC-101 and gemcitabine alone or combination for 48 h. Apoptotic cells were analyzed by flow cytometry. B. Western blot analysis showed CUDC-101 increased inhibition of gemcitabine on HDAC1, HDAC3, HDAC4 and p-EGFR proteins. Cyclin B1/CDK1 protein expression in CUDC-101 group was decreased. Acetylased histone H3 protein expression in CUDC-101 group and co-treatment group. C. Protein levels of p-p53, cl-caspase3 and the ratio of bax/bcl-2 were markedly increased in co-treatment group. Statistical significance was determined by a standard test (*P

    Techniques Used: Staining, Flow Cytometry, Cytometry, Western Blot, Inhibition, Expressing

    26) Product Images from "The Stemness-High Human Colorectal Cancer Cells Promote Angiogenesis by Producing Higher Amounts of Angiogenic Cytokines via Activation of the Egfr/Akt/Nf-κB Pathway"

    Article Title: The Stemness-High Human Colorectal Cancer Cells Promote Angiogenesis by Producing Higher Amounts of Angiogenic Cytokines via Activation of the Egfr/Akt/Nf-κB Pathway

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms22031355

    The EGFR/AKT pathway is responsible for the activation of NF-κB in the GATA6-overexpressing human CRC clones. ( A ) Total lysates (20 µg) prepared from three clones derived respectively from HCT-116 and HT-29 cells were subjected to western blot analyses using primary antibodies phospho-EGFR (Y1068), total EGFR, phospho-AKT (S473), and total AKT as probes, respectively. The quantitative results (bar graphs) obtained by densitometry are the mean ± SD of three independent experiments. * p
    Figure Legend Snippet: The EGFR/AKT pathway is responsible for the activation of NF-κB in the GATA6-overexpressing human CRC clones. ( A ) Total lysates (20 µg) prepared from three clones derived respectively from HCT-116 and HT-29 cells were subjected to western blot analyses using primary antibodies phospho-EGFR (Y1068), total EGFR, phospho-AKT (S473), and total AKT as probes, respectively. The quantitative results (bar graphs) obtained by densitometry are the mean ± SD of three independent experiments. * p

    Techniques Used: Activation Assay, Clone Assay, Derivative Assay, Western Blot

    27) Product Images from "MicroRNA-374a Promotes Hepatocellular Carcinoma Cell Proliferation by Targeting Mitogen-Inducible Gene 6 (MIG-6)"

    Article Title: MicroRNA-374a Promotes Hepatocellular Carcinoma Cell Proliferation by Targeting Mitogen-Inducible Gene 6 (MIG-6)

    Journal: Oncology Research

    doi: 10.3727/096504017X15000784459799

    miR-374a activates epidermal growth factor receptor (EGFR) and phosphoinositol 3-kinase/protein kinase B (AKT)/extracellular signal-regulated kinase (ERK) signaling pathways by regulation of MIG-6 in HepG2 cells. miR-374a mimic, pc-MIG-6, and their corresponding controls were transfected into HepG2 cells. (A) The protein levels of p-EGFR, EGFR, p-AKT, and p-ERK1/2 were measured by Western blot. (B) The mRNA expressions of p-EGFR, EGFR, p-AKT, and p-ERK1/2 were determined by RT-PCR. * p
    Figure Legend Snippet: miR-374a activates epidermal growth factor receptor (EGFR) and phosphoinositol 3-kinase/protein kinase B (AKT)/extracellular signal-regulated kinase (ERK) signaling pathways by regulation of MIG-6 in HepG2 cells. miR-374a mimic, pc-MIG-6, and their corresponding controls were transfected into HepG2 cells. (A) The protein levels of p-EGFR, EGFR, p-AKT, and p-ERK1/2 were measured by Western blot. (B) The mRNA expressions of p-EGFR, EGFR, p-AKT, and p-ERK1/2 were determined by RT-PCR. * p

    Techniques Used: Transfection, Western Blot, Reverse Transcription Polymerase Chain Reaction

    28) Product Images from "Novel matrine derivative MD-1 attenuates hepatic fibrosis by inhibiting EGFR activation of hepatic stellate cells"

    Article Title: Novel matrine derivative MD-1 attenuates hepatic fibrosis by inhibiting EGFR activation of hepatic stellate cells

    Journal: Protein & Cell

    doi: 10.1007/s13238-016-0285-2

    Inhibitory effects of MD-1 on the EGFR signaling pathway in HSC-T6 cells . (A) Immunostaining of EGFR-positive HSC-T6 cells showed MD-1 localization. Nuclei are stained with DAPI. Scale bar: 10 µm. (B) Similar to (A), but in EGFR-knockdown HSC-T6 cells. Scale bar: 10 µm. (C) Co-immunoprecipitation blots showed MD-1-Biotin was precipitated by EGFR in the MD-1-treated HSC-T6 cells, but not in the parental cells. (D) Representative Western blots to show the expression of EGFR, AKT, cyclin D1, and p-Smad. GAPDH was used as the loading control. Densitometry was performed to determine the relative expression levels normalized to GAPDH. * P
    Figure Legend Snippet: Inhibitory effects of MD-1 on the EGFR signaling pathway in HSC-T6 cells . (A) Immunostaining of EGFR-positive HSC-T6 cells showed MD-1 localization. Nuclei are stained with DAPI. Scale bar: 10 µm. (B) Similar to (A), but in EGFR-knockdown HSC-T6 cells. Scale bar: 10 µm. (C) Co-immunoprecipitation blots showed MD-1-Biotin was precipitated by EGFR in the MD-1-treated HSC-T6 cells, but not in the parental cells. (D) Representative Western blots to show the expression of EGFR, AKT, cyclin D1, and p-Smad. GAPDH was used as the loading control. Densitometry was performed to determine the relative expression levels normalized to GAPDH. * P

    Techniques Used: Immunostaining, Staining, Immunoprecipitation, Western Blot, Expressing

    Inhibitory effects of MD-1 on DMN-induced hepatic fibrosis in rats . (A) The sectioned rat liver tissues were stained with Van Gieson staining reagent to show the proliferated collagen fibers (stained with red) segmented and surrounded pseudolobules in the positive control group, and MD-1 treatment significantly reduced the collagen fibers. Original magnification: 200×. (B) Top: Expression of p-EGFR, p-AKT, α-SMA, cyclin D1, and p-Smad in sectioned rat liver was examined by immunohistochemistry. The percentages of positive cells (stained with brown) were counted within five fields of view for each section using a 20× objective lens. Original magnification: 100×. Bottom: Statistics for p-EGFR, p-AKT, α-SMA, cyclin D1, and p-Smad. * P
    Figure Legend Snippet: Inhibitory effects of MD-1 on DMN-induced hepatic fibrosis in rats . (A) The sectioned rat liver tissues were stained with Van Gieson staining reagent to show the proliferated collagen fibers (stained with red) segmented and surrounded pseudolobules in the positive control group, and MD-1 treatment significantly reduced the collagen fibers. Original magnification: 200×. (B) Top: Expression of p-EGFR, p-AKT, α-SMA, cyclin D1, and p-Smad in sectioned rat liver was examined by immunohistochemistry. The percentages of positive cells (stained with brown) were counted within five fields of view for each section using a 20× objective lens. Original magnification: 100×. Bottom: Statistics for p-EGFR, p-AKT, α-SMA, cyclin D1, and p-Smad. * P

    Techniques Used: Staining, Positive Control, Expressing, Immunohistochemistry

    29) Product Images from "Genistein Increases Epidermal Growth Factor Receptor Signaling and Promotes Tumor Progression in Advanced Human Prostate Cancer"

    Article Title: Genistein Increases Epidermal Growth Factor Receptor Signaling and Promotes Tumor Progression in Advanced Human Prostate Cancer

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0020034

    Effects of genistein on EGFR signaling. a) Genistein increases phosphorylation of EGFR at tyrosine 1068 residue. Below is the quantification of band intensity for phosphorylated EGFR from protein lysates of 5 untreated control and 6 genistein-treated tumors. Columns : mean ratio of phosphorylated EGFR/total EGFR protein band intensity ± SD. b) Genistein affects phosphorylation of Src at tyrosine 416 residue. Activated Src expression is significantly higher in genistein-treated tumors than that in untreated control. Lanes represent proteins from individual tumors.
    Figure Legend Snippet: Effects of genistein on EGFR signaling. a) Genistein increases phosphorylation of EGFR at tyrosine 1068 residue. Below is the quantification of band intensity for phosphorylated EGFR from protein lysates of 5 untreated control and 6 genistein-treated tumors. Columns : mean ratio of phosphorylated EGFR/total EGFR protein band intensity ± SD. b) Genistein affects phosphorylation of Src at tyrosine 416 residue. Activated Src expression is significantly higher in genistein-treated tumors than that in untreated control. Lanes represent proteins from individual tumors.

    Techniques Used: Expressing

    30) Product Images from "PIM1 inhibitor synergizes the anti-tumor effect of osimertinib via STAT3 dephosphorylation in EGFR-mutant non-small cell lung cancer"

    Article Title: PIM1 inhibitor synergizes the anti-tumor effect of osimertinib via STAT3 dephosphorylation in EGFR-mutant non-small cell lung cancer

    Journal: Annals of Translational Medicine

    doi: 10.21037/atm.2020.02.43

    Effect of osimertinib on the expression of PIM1, caspase 3, and PARP, and the total levels or phosphorylation inhibition of STAT3, EGFR, AKT, and ERK. H1975 and PC9 cells were treated with vehicle, 5 µM CX-6258 HCl, 5 µM osimertinib, or a combination of both (5 µM) for 6 h following EGF stimulation, and the extracts were blotted using the indicated antibodies. EGFR, epidermal growth factor receptor.
    Figure Legend Snippet: Effect of osimertinib on the expression of PIM1, caspase 3, and PARP, and the total levels or phosphorylation inhibition of STAT3, EGFR, AKT, and ERK. H1975 and PC9 cells were treated with vehicle, 5 µM CX-6258 HCl, 5 µM osimertinib, or a combination of both (5 µM) for 6 h following EGF stimulation, and the extracts were blotted using the indicated antibodies. EGFR, epidermal growth factor receptor.

    Techniques Used: Expressing, Inhibition

    CX-6258 HCl combined with osimertinib inhibited the expression of PIM1, p-EGFR, and caspase-3 in the tumor tissue. Immunohistochemistry images for PIM1, p-EGFR, and caspase-3 staining of tumor tissue in BALB/c nu/nu mice. All images were 20× magnification; bar =50 µm.
    Figure Legend Snippet: CX-6258 HCl combined with osimertinib inhibited the expression of PIM1, p-EGFR, and caspase-3 in the tumor tissue. Immunohistochemistry images for PIM1, p-EGFR, and caspase-3 staining of tumor tissue in BALB/c nu/nu mice. All images were 20× magnification; bar =50 µm.

    Techniques Used: Expressing, Immunohistochemistry, Staining, Mouse Assay

    31) Product Images from "Multiple receptor tyrosine kinase activation attenuates therapeutic efficacy of the fibroblast growth factor receptor 2 inhibitor AZD4547 in FGFR2 amplified gastric cancer"

    Article Title: Multiple receptor tyrosine kinase activation attenuates therapeutic efficacy of the fibroblast growth factor receptor 2 inhibitor AZD4547 in FGFR2 amplified gastric cancer

    Journal: Oncotarget

    doi:

    FGFR2 positive tumors showed copositivity of enriched drug resistance related RTKs Representative immunohistochemical analysis of one patient with FGFR2 positive GC is shown. Copositivity of HER3, MET and EGFR were accompanied by FGFR2 positivity.
    Figure Legend Snippet: FGFR2 positive tumors showed copositivity of enriched drug resistance related RTKs Representative immunohistochemical analysis of one patient with FGFR2 positive GC is shown. Copositivity of HER3, MET and EGFR were accompanied by FGFR2 positivity.

    Techniques Used: Immunohistochemistry

    32) Product Images from "HCRP-1 regulates EGFR–AKT–BIM-mediated anoikis resistance and serves as a prognostic marker in human colon cancer"

    Article Title: HCRP-1 regulates EGFR–AKT–BIM-mediated anoikis resistance and serves as a prognostic marker in human colon cancer

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-1217-2

    Hypothetic model of HCRP-1 depletion inhibits BIM expression via the EGFR–AKT signaling pathway
    Figure Legend Snippet: Hypothetic model of HCRP-1 depletion inhibits BIM expression via the EGFR–AKT signaling pathway

    Techniques Used: Expressing

    Suppression of EGFR blocks the loss of HCRP-1 induced by the AKT signaling pathway, upregulates BIM expression, and suppresses anoikis resistance. a , b HCT116 and SW620 cells were pretreated with HCRP-1 siRNA and the EGFR inhibitor, AG1478 (5 mM/L), and then suspended in six-well plates with low-attachment surface. The whole-cell lysates were analyzed for the protein levels of p-EGFR, EGFR, p-AKT, AKT, BIM, and β-actin by using western blot. c , d Cell anoikis was detected by flow cytometry. HCT116 and SW620 cells were pretreated with HCRP-1 siRNA and the EGFR inhibitor, AG1478 (5 mM/L), and then suspended in six-well plates with low- attachment surface. Cells that did not adhere were subjected to flow cytometry detection. e and f HCT116 and SW620 cells were co-transfected with HCRP-1 siRNA and/or EGFR siRNA. Levels of p-EGFR, EGFR, p-AKT, AKT, BIM, and β-actin in cells that did not adhere were determined by Western blot. g , h Cell anoikis was detected by flow cytometry in colon cancer cells pretreated with HCRP-1 siRNA and/or EGFR siRNA. Error bars indicate mean ± SD. Every experiment was repeated at least three times. * P
    Figure Legend Snippet: Suppression of EGFR blocks the loss of HCRP-1 induced by the AKT signaling pathway, upregulates BIM expression, and suppresses anoikis resistance. a , b HCT116 and SW620 cells were pretreated with HCRP-1 siRNA and the EGFR inhibitor, AG1478 (5 mM/L), and then suspended in six-well plates with low-attachment surface. The whole-cell lysates were analyzed for the protein levels of p-EGFR, EGFR, p-AKT, AKT, BIM, and β-actin by using western blot. c , d Cell anoikis was detected by flow cytometry. HCT116 and SW620 cells were pretreated with HCRP-1 siRNA and the EGFR inhibitor, AG1478 (5 mM/L), and then suspended in six-well plates with low- attachment surface. Cells that did not adhere were subjected to flow cytometry detection. e and f HCT116 and SW620 cells were co-transfected with HCRP-1 siRNA and/or EGFR siRNA. Levels of p-EGFR, EGFR, p-AKT, AKT, BIM, and β-actin in cells that did not adhere were determined by Western blot. g , h Cell anoikis was detected by flow cytometry in colon cancer cells pretreated with HCRP-1 siRNA and/or EGFR siRNA. Error bars indicate mean ± SD. Every experiment was repeated at least three times. * P

    Techniques Used: Expressing, Western Blot, Flow Cytometry, Cytometry, Transfection

    Inhibition of HCRP-1 activates the EGFR/AKT signaling pathway and suppresses BIM protein expression. a , b HCT116 and SW620 cells were transfected with si-HCRP-1 or control siRNA, and then cells were harvested and submitted to Western blot detection for the protein expression of BIM, FoxO3a, EGFR, AKT, ERK, Mcl-1, Bcl-2, and β-actin. Phosphorylated forms of EGFR, AKT, FoxO3a, and ERK were also detected by western blot with the corresponding antibodies. c–f Cells were treatedwith CHX for 0, 0.5 , 1, and 2 h after transfection with si-HCRP-1 or control siRNA for 48 h, lysates obtained from these cells were submitted to western blot detection for the protein expression of EGFR. Error bars indicate mean ± SD. Every experiment was repeated at least three times. * P
    Figure Legend Snippet: Inhibition of HCRP-1 activates the EGFR/AKT signaling pathway and suppresses BIM protein expression. a , b HCT116 and SW620 cells were transfected with si-HCRP-1 or control siRNA, and then cells were harvested and submitted to Western blot detection for the protein expression of BIM, FoxO3a, EGFR, AKT, ERK, Mcl-1, Bcl-2, and β-actin. Phosphorylated forms of EGFR, AKT, FoxO3a, and ERK were also detected by western blot with the corresponding antibodies. c–f Cells were treatedwith CHX for 0, 0.5 , 1, and 2 h after transfection with si-HCRP-1 or control siRNA for 48 h, lysates obtained from these cells were submitted to western blot detection for the protein expression of EGFR. Error bars indicate mean ± SD. Every experiment was repeated at least three times. * P

    Techniques Used: Inhibition, Expressing, Transfection, Western Blot

    33) Product Images from "IKKα Promotes the Progression and Metastasis of Non-Small Cell Lung Cancer Independently of its Subcellular Localization"

    Article Title: IKKα Promotes the Progression and Metastasis of Non-Small Cell Lung Cancer Independently of its Subcellular Localization

    Journal: Computational and Structural Biotechnology Journal

    doi: 10.1016/j.csbj.2019.02.003

    Molecular characterization of the different isolated clones that overexpress IKKα in the cytoplasm or in the nucleus of the H460 cells. (A) WB of total protein extracts showing the hybridization of different C-H460-IKKα pools with the indicated antibodies. Note that those clones with increased levels of IKKα (C-1, C-6, C-7 and C-12) exhibit overactivation of the classical NF-κB pathway (elevated levels of P-p65 and P-IκBα); as well as activation of EGFR (measured as P-EGFR). They also present increased levels of Cyclin D1 expression with respect to the values observed in the Control-H460 cells and those of the clones that do not express the transgene (C-8, C-9 and C-2). (B) Levels of P-p65 in the N-H460-IKKα cells are similar to those of Control cells, indicating that no enhanced activation of the NF-κB is observed when IKKα is expressed in the nucleus of the H460 cells. Enhanced P-Smad2/3 is also detected in the N-H460-IKKα mice, while it is not enhanced in the C-H460-IKKα cells (A). (C) WB showing overactivation of EGFR in the C-H460-IKKα cells, but not in the N-H460-IKKα ones. (D) Increased activation of Erk1/2 is observed in both types of cells overexpressing IKKα. (E) Levels of Cyclin D1 are increased in the N-H460-IKKα clones regarding the values in the Control-H460 cells. (F) Overactivation of mTor (increased levels of P-mTor) is detected in both C-and-N-H460-IKKα cells. (G) N-H460-IKKα cells present increased levels of P-c-Myc and Snail. Increased levels of P-p38 and Podoplanin are observed in the C-and-N-H460-IKKα cells. (H) Levels of IKKα expression in the N-H460-IKKα pools N-2 and N-10 are shown. Tubulin and GAPDH are used as loading control. Different C– H460-IKKα, N-H460-IKKα and Control-H460 pools are shown.
    Figure Legend Snippet: Molecular characterization of the different isolated clones that overexpress IKKα in the cytoplasm or in the nucleus of the H460 cells. (A) WB of total protein extracts showing the hybridization of different C-H460-IKKα pools with the indicated antibodies. Note that those clones with increased levels of IKKα (C-1, C-6, C-7 and C-12) exhibit overactivation of the classical NF-κB pathway (elevated levels of P-p65 and P-IκBα); as well as activation of EGFR (measured as P-EGFR). They also present increased levels of Cyclin D1 expression with respect to the values observed in the Control-H460 cells and those of the clones that do not express the transgene (C-8, C-9 and C-2). (B) Levels of P-p65 in the N-H460-IKKα cells are similar to those of Control cells, indicating that no enhanced activation of the NF-κB is observed when IKKα is expressed in the nucleus of the H460 cells. Enhanced P-Smad2/3 is also detected in the N-H460-IKKα mice, while it is not enhanced in the C-H460-IKKα cells (A). (C) WB showing overactivation of EGFR in the C-H460-IKKα cells, but not in the N-H460-IKKα ones. (D) Increased activation of Erk1/2 is observed in both types of cells overexpressing IKKα. (E) Levels of Cyclin D1 are increased in the N-H460-IKKα clones regarding the values in the Control-H460 cells. (F) Overactivation of mTor (increased levels of P-mTor) is detected in both C-and-N-H460-IKKα cells. (G) N-H460-IKKα cells present increased levels of P-c-Myc and Snail. Increased levels of P-p38 and Podoplanin are observed in the C-and-N-H460-IKKα cells. (H) Levels of IKKα expression in the N-H460-IKKα pools N-2 and N-10 are shown. Tubulin and GAPDH are used as loading control. Different C– H460-IKKα, N-H460-IKKα and Control-H460 pools are shown.

    Techniques Used: Isolation, Clone Assay, Western Blot, Hybridization, Activation Assay, Expressing, Mouse Assay

    34) Product Images from "Pharmacological targeting of BET proteins inhibits renal fibroblast activation and alleviates renal fibrosis"

    Article Title: Pharmacological targeting of BET proteins inhibits renal fibroblast activation and alleviates renal fibrosis

    Journal: Oncotarget

    doi: 10.18632/oncotarget.12498

    I-BET151 inhibits phosphorylation of EGFR and PDGFRβ in obstructed kidneys A. Kidney tissue lysates were subjected to immunoblot analysis with antibodies against phospho-EGFR (Tyr1068), phospho-PDGFRβ(Tyr751), EGFR, PDGFRβ,and GAPDH. All of these proteins were quantified by densitometry, andphospho-EGFR and PDGFRβ B. and D. were normalized to their total protein levels; EGFR C. and PDGFR E. were normalized with Tubulin. Data are represented as the mean ± SEM. Means with different superscript letters (a-c) are significantly different from one another ( P
    Figure Legend Snippet: I-BET151 inhibits phosphorylation of EGFR and PDGFRβ in obstructed kidneys A. Kidney tissue lysates were subjected to immunoblot analysis with antibodies against phospho-EGFR (Tyr1068), phospho-PDGFRβ(Tyr751), EGFR, PDGFRβ,and GAPDH. All of these proteins were quantified by densitometry, andphospho-EGFR and PDGFRβ B. and D. were normalized to their total protein levels; EGFR C. and PDGFR E. were normalized with Tubulin. Data are represented as the mean ± SEM. Means with different superscript letters (a-c) are significantly different from one another ( P

    Techniques Used:

    35) Product Images from "Histone deacetylase 6 inhibition counteracts the epithelial–mesenchymal transition of peritoneal mesothelial cells and prevents peritoneal fibrosis"

    Article Title: Histone deacetylase 6 inhibition counteracts the epithelial–mesenchymal transition of peritoneal mesothelial cells and prevents peritoneal fibrosis

    Journal: Oncotarget

    doi: 10.18632/oncotarget.20982

    Inhibition of HDAC6 abrogates the activation of EGFR/STAT3 signaling pathway in the peritoneum exposed to high glucose dialysate Peritoneal membrane was collected at 28 days after PDF injection with or without administration of TA (70 mg/kg, daily). (A) The peritoneal tissue lysates were subjected to immunoblot analysis with specific antibodies against p-EGFR, EGFR, p-STAT3, STAT3 or GAPDH. (B) Expression level of p-EGFR was quantified by densitometry and normalized with total EGFR. (D) Expression level of p-STAT3 was quantified by densitometry and normalized with total STAT3. Expression levels of total EGFR (C) or total STAT3 (E) were quantified by densitometry and normalized with total GAPDH. Data are represented as the means±SD (n=6). Means with different letters are significantly different from one another (P
    Figure Legend Snippet: Inhibition of HDAC6 abrogates the activation of EGFR/STAT3 signaling pathway in the peritoneum exposed to high glucose dialysate Peritoneal membrane was collected at 28 days after PDF injection with or without administration of TA (70 mg/kg, daily). (A) The peritoneal tissue lysates were subjected to immunoblot analysis with specific antibodies against p-EGFR, EGFR, p-STAT3, STAT3 or GAPDH. (B) Expression level of p-EGFR was quantified by densitometry and normalized with total EGFR. (D) Expression level of p-STAT3 was quantified by densitometry and normalized with total STAT3. Expression levels of total EGFR (C) or total STAT3 (E) were quantified by densitometry and normalized with total GAPDH. Data are represented as the means±SD (n=6). Means with different letters are significantly different from one another (P

    Techniques Used: Inhibition, Activation Assay, Injection, Expressing

    HDAC6 is required for phosphorylation of EGFR and STAT3 in peritoneal mesothelial cells exposed to TGF-β1 Serum-starved HPMCs were pretreated with various concentrations of TA (0-20 μM) for 1 hour (A) or transfected with siRNA targeting HDAC6 or scrambled (Con) siRNA for 24 hours (E) and then exposed to TGF-β1 (10 ng/ml) for an additional 24h. (A, E) Cell lysates were subjected to immunoblot analysis with antibodies p-EGFR, EGFR, p-STAT3, STAT3 or β-actin. (B, F) Expression level of p-EGFR was quantified by densitometry and normalized with EGFR. (C, G) Expression level of total EGFR was quantified by densitometry and normalized with β-actin. (D, H) Expression level of p-STAT3 was quantified by densitometry and normalized with total STAT3. Values are means±SD of at least three independent experiments. Bars with different letters (a-d) for each molecule are significantly different from one another (P
    Figure Legend Snippet: HDAC6 is required for phosphorylation of EGFR and STAT3 in peritoneal mesothelial cells exposed to TGF-β1 Serum-starved HPMCs were pretreated with various concentrations of TA (0-20 μM) for 1 hour (A) or transfected with siRNA targeting HDAC6 or scrambled (Con) siRNA for 24 hours (E) and then exposed to TGF-β1 (10 ng/ml) for an additional 24h. (A, E) Cell lysates were subjected to immunoblot analysis with antibodies p-EGFR, EGFR, p-STAT3, STAT3 or β-actin. (B, F) Expression level of p-EGFR was quantified by densitometry and normalized with EGFR. (C, G) Expression level of total EGFR was quantified by densitometry and normalized with β-actin. (D, H) Expression level of p-STAT3 was quantified by densitometry and normalized with total STAT3. Values are means±SD of at least three independent experiments. Bars with different letters (a-d) for each molecule are significantly different from one another (P

    Techniques Used: Transfection, Expressing

    36) Product Images from "Overcoming drug-tolerant cancer cell subpopulations showing AXL activation and epithelial–mesenchymal transition is critical in conquering ALK-positive lung cancer"

    Article Title: Overcoming drug-tolerant cancer cell subpopulations showing AXL activation and epithelial–mesenchymal transition is critical in conquering ALK-positive lung cancer

    Journal: Oncotarget

    doi: 10.18632/oncotarget.25531

    Establishment of ALK-TKI–resistant H2228 cells (A) H2228-CRR, H2228-ALR and H2228-CER cells are resistant to crizotinib, alectinib and ceritinib, respectively. The results of cell viability assays are shown. Data, mean ± SEM from three independent experiments. (B) FISH analysis shows a decrease of the ALK fusion gene in ALK-TKI–resistant H2228 cells compared with H2228 cells (red, ALK3’ ; green, ALK5’ ). The percentage of ALK-FISH positive cells are shown in the lower right corners. (C) Protein levels of p-ALK, ALK, p-EGFR, EGFR, p-AKT, AKT, p-ERK and ERK were analyzed by western blotting. ALK-TKI–resistant cells showed markedly decreased p-ALK and ALK expressions.
    Figure Legend Snippet: Establishment of ALK-TKI–resistant H2228 cells (A) H2228-CRR, H2228-ALR and H2228-CER cells are resistant to crizotinib, alectinib and ceritinib, respectively. The results of cell viability assays are shown. Data, mean ± SEM from three independent experiments. (B) FISH analysis shows a decrease of the ALK fusion gene in ALK-TKI–resistant H2228 cells compared with H2228 cells (red, ALK3’ ; green, ALK5’ ). The percentage of ALK-FISH positive cells are shown in the lower right corners. (C) Protein levels of p-ALK, ALK, p-EGFR, EGFR, p-AKT, AKT, p-ERK and ERK were analyzed by western blotting. ALK-TKI–resistant cells showed markedly decreased p-ALK and ALK expressions.

    Techniques Used: Fluorescence In Situ Hybridization, Western Blot

    Related Articles

    Western Blot:

    Article Title: A gain-of-function mutant p53-HSF1 feed forward circuit governs adaptation of cancer cells to proteotoxic stress
    Article Snippet: .. Immunoblots and immunoprecipitations For immunoblots, equal total protein of cell lysates (2.5–20 μ g) were detected with antibodies to mouse p53 (FL393), human p53 (PAb1801; Santa Cruz Biotechnology), HSF1, p-Ser326 HSF1, AKT, pAKT, Erk, pErk, Hsp70, Hsp90, Hsp90α , EGFR, EGRF-Tyr845P (all Cell Signaling, Danvers, MA, USA), ErbB2, actin, GAPDH, GTS, GFP, HSc70 and HDAC1 (all Neomarkers, Fremont, CA, USA)., , SDS-PAAG gels (6%) were used to detect slower migrating HS-activated HSF1 used in experiments presented in and . ..

    Immunohistochemistry:

    Article Title: Elevated Expression of Fn14 in Non-Small Cell Lung Cancer Correlates with Activated EGFR and Promotes Tumor Cell Migration and Invasion
    Article Snippet: .. IHC analysis for Fn14 was performed using the Fn14 monoclonal antibody P4A8 (Biogen Idec, Inc., Weston, MA), as previously described. p-EGFR analysis was performed using an antibody specific for EGFR-Y1068 (Cell Signaling Technologies, Beverly, CA). .. Human NSCLC cell lines H520, H2122, A549, H1703, H358, H3255, H1975, HCC2279, and HCC827 (ATCC, Manassas, VA) were maintained in RPMI 1640 medium (Invitrogen, Carlsbad, CA), supplemented with 10% heat-inactivated fetal bovine serum (FBS) in a 37°C, 5% CO2 atmosphere.

    Synthesized:

    Article Title: Inhibition of histone deacetylases sensitizes EGF receptor‐TK inhibitor‐resistant non‐small‐cell lung cancer cells to erlotinib in vitro and in vivo) Inhibition of histone deacetylases sensitizes EGF receptor‐TK inhibitor‐resistant non‐small‐cell lung cancer cells to erlotinib in vitro and in vivo
    Article Snippet: Erlotinib and SAHA were purchased from Selleck Chemicals (Houston, TX, USA). .. YF454A [N1‐((5‐(5‐pyrimidinyl)‐2‐thiopheneyl) methyl)‐N7‐hydroxyN1‐(4‐methoxyphenyl) heptane‐diamide] and other lead compounds were synthesized in house (Yang et al., (Ser473 (Thr202/ Tyr204 ), ERK1/2, p‐EGFR (pTyr1068 ), EGFR, p‐Her2 (Tyr1221/1222 ), Her2, p‐c‐Met(Tyr1234/1235 ), c‐Met, p‐IGF1R (pTyr1316 , cyclin D1 and E2F1 antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). .. Antibodies against Cdc25A, p‐Rb (Ser807/811 ) and Rb were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

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    Differential expression of <t>EGFR,</t> p-EGFR and <t>PKC-α</t> between MEKi-Se and MEKi-Re LGSC cell lines by WB. a Confirmation of the RPPA results in untreated MEKi-Se and MEKi-Re lines ( a , discovery cohort). EGFR, p-EGFR and PKC-α were increased in MEKi-Re lines (n = 5) compared to MEKi-Se lines (n = 2). b Validation of these protein biomarkers in three newly established LGSC cell lines classified according to their MEKi responsiveness (validation cohort). As found in the cell lines analyzed by RPPA, the new MEKi-Re line (n = 1) expressed higher levels of EGFR, p-EGFR and PKC-α compared to the two new MEKi-Se lines tested (n = 2). c , d Confirmation of RPPA results in MEKi treated cell lines. With MEKi treatment (selumetinib 1 μM and trametinib 0.1 μM) p-EGFR expression remained higher in MEKi-Re lines. As previously described, trametinib showed stronger inhibitory effects on MAPK (p-MAPK or p-ERK1/2) than selumetinib, even when used at ten times lower dose. (*) No DMSO. (**) Untreated VOA-4627 cells to control for drug inhibition effects on MAPK pathway
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    Differential expression of EGFR, p-EGFR and PKC-α between MEKi-Se and MEKi-Re LGSC cell lines by WB. a Confirmation of the RPPA results in untreated MEKi-Se and MEKi-Re lines ( a , discovery cohort). EGFR, p-EGFR and PKC-α were increased in MEKi-Re lines (n = 5) compared to MEKi-Se lines (n = 2). b Validation of these protein biomarkers in three newly established LGSC cell lines classified according to their MEKi responsiveness (validation cohort). As found in the cell lines analyzed by RPPA, the new MEKi-Re line (n = 1) expressed higher levels of EGFR, p-EGFR and PKC-α compared to the two new MEKi-Se lines tested (n = 2). c , d Confirmation of RPPA results in MEKi treated cell lines. With MEKi treatment (selumetinib 1 μM and trametinib 0.1 μM) p-EGFR expression remained higher in MEKi-Re lines. As previously described, trametinib showed stronger inhibitory effects on MAPK (p-MAPK or p-ERK1/2) than selumetinib, even when used at ten times lower dose. (*) No DMSO. (**) Untreated VOA-4627 cells to control for drug inhibition effects on MAPK pathway

    Journal: Cancer Cell International

    Article Title: Markers of MEK inhibitor resistance in low-grade serous ovarian cancer: EGFR is a potential therapeutic target

    doi: 10.1186/s12935-019-0725-1

    Figure Lengend Snippet: Differential expression of EGFR, p-EGFR and PKC-α between MEKi-Se and MEKi-Re LGSC cell lines by WB. a Confirmation of the RPPA results in untreated MEKi-Se and MEKi-Re lines ( a , discovery cohort). EGFR, p-EGFR and PKC-α were increased in MEKi-Re lines (n = 5) compared to MEKi-Se lines (n = 2). b Validation of these protein biomarkers in three newly established LGSC cell lines classified according to their MEKi responsiveness (validation cohort). As found in the cell lines analyzed by RPPA, the new MEKi-Re line (n = 1) expressed higher levels of EGFR, p-EGFR and PKC-α compared to the two new MEKi-Se lines tested (n = 2). c , d Confirmation of RPPA results in MEKi treated cell lines. With MEKi treatment (selumetinib 1 μM and trametinib 0.1 μM) p-EGFR expression remained higher in MEKi-Re lines. As previously described, trametinib showed stronger inhibitory effects on MAPK (p-MAPK or p-ERK1/2) than selumetinib, even when used at ten times lower dose. (*) No DMSO. (**) Untreated VOA-4627 cells to control for drug inhibition effects on MAPK pathway

    Article Snippet: Western blot analysis Cell lysates were prepared according to previously published protocols [ ], then 20 μg samples were separated on an 8% SDS-PAGE gel, transferred to nitrocellulose membranes and probed with primary antibodies including ERK1/2 (Millipore, Cat. No. 06-182), p-MAPK (p-ERK1/2, Cell Signaling, Cat. No. 4376S), MEK1/2 (Cell Signaling, Cat. No. 9122), p-MEK1/2 (Cell Signaling, Cat. No. 9154), PKC-alpha (Cell Signaling, Cat. No. 2056), EGFR (Santa Cruz, Cat. No. 71032), p-EGFR (Cell Signaling, Cat. No. 2234), PARP (Cell Signaling, Cat. No. 9542), and c-PARP (Cell Signaling, Cat. No. 9541S).

    Techniques: Expressing, Western Blot, Inhibition

    Overexpression of Cx32 promotes the proliferation of SMMC-7721 cells and protects cells from SN-induced apoptosis in vivo. a. Representative images of the nude mouse xenograft model. b. Tumor growth curves. Overexpression of Cx32 promoted the tumor growth in nude mice and significantly reduced the growth suppression mediated by intragastric injection of SN (0.5 mg/kg). c. Representative images of tumors from the sacrificed nude mice. d. Representative images of IHC for Cx32, EGFR and Src in tumors generated from SMMC-Vector and SMMC-Cx32 cells. Scale bars: 50 μm. e. Overexpression of Cx32 inhibited the SN-induced increase in the levels of cleaved-caspase3 by increasing the levels of EGFR and Src. ## , P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Cx32 exerts anti-apoptotic and pro-tumor effects via the epidermal growth factor receptor pathway in hepatocellular carcinoma

    doi: 10.1186/s13046-019-1142-y

    Figure Lengend Snippet: Overexpression of Cx32 promotes the proliferation of SMMC-7721 cells and protects cells from SN-induced apoptosis in vivo. a. Representative images of the nude mouse xenograft model. b. Tumor growth curves. Overexpression of Cx32 promoted the tumor growth in nude mice and significantly reduced the growth suppression mediated by intragastric injection of SN (0.5 mg/kg). c. Representative images of tumors from the sacrificed nude mice. d. Representative images of IHC for Cx32, EGFR and Src in tumors generated from SMMC-Vector and SMMC-Cx32 cells. Scale bars: 50 μm. e. Overexpression of Cx32 inhibited the SN-induced increase in the levels of cleaved-caspase3 by increasing the levels of EGFR and Src. ## , P

    Article Snippet: EGFR, p-EGFR (Tyr845), PARP, cleaved-Caspase3, Erk 1/2, p-Erk 1/2 (Thr202/Tyr204), STAT3, p-STAT3 (Tyr705), Bcl-2, Bak, Bax, and Src primary antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Over Expression, In Vivo, Mouse Assay, Injection, Immunohistochemistry, Generated, Plasmid Preparation

    APE1 mediates bile salts-induced STAT3 activation via an EGFR-dependent mechanism Immunoblot analysis of CPB ( A ), FLO-1 ( B ) and OE33 ( C ) cells pretreated with EGFR inhibitor (Gefitinib, 25 μM) followed by exposure to acidic (pH4) bile salts (100 μM). The samples were analyzed for the indicated proteins, β-actin was used as an internal control. ( D ) Immunoblot analysis of OE33 cells with EGFR-knockdown via EGFR siRNA followed by treatment with acidic (pH4) bile salts (100 μM) for 20 minutes and allowed to recover in complete media. The samples were collected at 3 and 6h post recovery and analyzed for the indicated proteins, β-actin was used as an internal control. Immunoprecipitation (IP) of APE1 ( E ), EGFR ( F ) and STAT3 ( G ) in OE33 cells treated with acidic (pH4) bile salts (100 μM) and immunoblotted for the indicated proteins. Results shown are representative of at least three independent experiments.

    Journal: Oncogene

    Article Title: Exposure of Barrett’s and esophageal adenocarcinoma cells to Bile acids activates EGFR-STAT3 signaling axis via induction of APE1

    doi: 10.1038/s41388-018-0388-8

    Figure Lengend Snippet: APE1 mediates bile salts-induced STAT3 activation via an EGFR-dependent mechanism Immunoblot analysis of CPB ( A ), FLO-1 ( B ) and OE33 ( C ) cells pretreated with EGFR inhibitor (Gefitinib, 25 μM) followed by exposure to acidic (pH4) bile salts (100 μM). The samples were analyzed for the indicated proteins, β-actin was used as an internal control. ( D ) Immunoblot analysis of OE33 cells with EGFR-knockdown via EGFR siRNA followed by treatment with acidic (pH4) bile salts (100 μM) for 20 minutes and allowed to recover in complete media. The samples were collected at 3 and 6h post recovery and analyzed for the indicated proteins, β-actin was used as an internal control. Immunoprecipitation (IP) of APE1 ( E ), EGFR ( F ) and STAT3 ( G ) in OE33 cells treated with acidic (pH4) bile salts (100 μM) and immunoblotted for the indicated proteins. Results shown are representative of at least three independent experiments.

    Article Snippet: Antibodies against p-STAT3 (Y705), STAT3, p-EGFR (Y1068), EGFR, BCL-xL, Survivin, α – Tubulin were purchased from Cell Signaling Technology (Danvers, Massachusetts, USA).

    Techniques: Activation Assay, Immunoprecipitation

    APE1 co-localizes with EGFR and STAT3 (A and B) In situ proximity ligation assay (PLA) demonstrates the interaction of p-EGFR and p-STAT3 with APE1. (C) In situ proximity ligation assay (PLA) demonstrates the interaction of p-EGFR and p-STAT3. Protein interactions (red fluorescent signals) were revealed by PLA anti-rabbit plus probe and PLA anti-mouse minus probe in OE33 cells treated with acidic (pH4) bile salts (100 μM) as mentioned in the materials and methods. Nuclei were stained with DAPI (blue). Results shown are representative of at least three independent experiments.

    Journal: Oncogene

    Article Title: Exposure of Barrett’s and esophageal adenocarcinoma cells to Bile acids activates EGFR-STAT3 signaling axis via induction of APE1

    doi: 10.1038/s41388-018-0388-8

    Figure Lengend Snippet: APE1 co-localizes with EGFR and STAT3 (A and B) In situ proximity ligation assay (PLA) demonstrates the interaction of p-EGFR and p-STAT3 with APE1. (C) In situ proximity ligation assay (PLA) demonstrates the interaction of p-EGFR and p-STAT3. Protein interactions (red fluorescent signals) were revealed by PLA anti-rabbit plus probe and PLA anti-mouse minus probe in OE33 cells treated with acidic (pH4) bile salts (100 μM) as mentioned in the materials and methods. Nuclei were stained with DAPI (blue). Results shown are representative of at least three independent experiments.

    Article Snippet: Antibodies against p-STAT3 (Y705), STAT3, p-EGFR (Y1068), EGFR, BCL-xL, Survivin, α – Tubulin were purchased from Cell Signaling Technology (Danvers, Massachusetts, USA).

    Techniques: In Situ, Proximity Ligation Assay, Staining

    Ectopic expression of wild-type (WT) EGFR, activated EGFR mutants, and the K-ras V12 mutant in rat lung epithelial cells (RL-65) induces Fn14 expression. Total cellular lysates from RL-65 cells stably expressing the indicated EGFR receptors or K-ras V12

    Journal: The American Journal of Pathology

    Article Title: Elevated Expression of Fn14 in Non-Small Cell Lung Cancer Correlates with Activated EGFR and Promotes Tumor Cell Migration and Invasion

    doi: 10.1016/j.ajpath.2012.03.026

    Figure Lengend Snippet: Ectopic expression of wild-type (WT) EGFR, activated EGFR mutants, and the K-ras V12 mutant in rat lung epithelial cells (RL-65) induces Fn14 expression. Total cellular lysates from RL-65 cells stably expressing the indicated EGFR receptors or K-ras V12

    Article Snippet: IHC analysis for Fn14 was performed using the Fn14 monoclonal antibody P4A8 (Biogen Idec, Inc., Weston, MA), as previously described. p-EGFR analysis was performed using an antibody specific for EGFR-Y1068 (Cell Signaling Technologies, Beverly, CA).

    Techniques: Expressing, Mutagenesis, Stable Transfection

    Erlotinib treatment of TKI-sensitive NSCLC cells decreases Fn14 expression. A: Serum-starved HCC827 cells containing the EGFR E746-A750–activating mutation were treated with vehicle for 12 hours or 1 μmol/L erlotinib for the indicated

    Journal: The American Journal of Pathology

    Article Title: Elevated Expression of Fn14 in Non-Small Cell Lung Cancer Correlates with Activated EGFR and Promotes Tumor Cell Migration and Invasion

    doi: 10.1016/j.ajpath.2012.03.026

    Figure Lengend Snippet: Erlotinib treatment of TKI-sensitive NSCLC cells decreases Fn14 expression. A: Serum-starved HCC827 cells containing the EGFR E746-A750–activating mutation were treated with vehicle for 12 hours or 1 μmol/L erlotinib for the indicated

    Article Snippet: IHC analysis for Fn14 was performed using the Fn14 monoclonal antibody P4A8 (Biogen Idec, Inc., Weston, MA), as previously described. p-EGFR analysis was performed using an antibody specific for EGFR-Y1068 (Cell Signaling Technologies, Beverly, CA).

    Techniques: Expressing, Mutagenesis

    Fn14 expression in human NSCLC specimens and correlation with EGFR phosphorylation. A: Fn14 and p-EGFR staining on representative samples from two patients with lung adenocarcinoma (×5 objective, Aperio GL Scanner; Aperio, Vista, CA). Insets:

    Journal: The American Journal of Pathology

    Article Title: Elevated Expression of Fn14 in Non-Small Cell Lung Cancer Correlates with Activated EGFR and Promotes Tumor Cell Migration and Invasion

    doi: 10.1016/j.ajpath.2012.03.026

    Figure Lengend Snippet: Fn14 expression in human NSCLC specimens and correlation with EGFR phosphorylation. A: Fn14 and p-EGFR staining on representative samples from two patients with lung adenocarcinoma (×5 objective, Aperio GL Scanner; Aperio, Vista, CA). Insets:

    Article Snippet: IHC analysis for Fn14 was performed using the Fn14 monoclonal antibody P4A8 (Biogen Idec, Inc., Weston, MA), as previously described. p-EGFR analysis was performed using an antibody specific for EGFR-Y1068 (Cell Signaling Technologies, Beverly, CA).

    Techniques: Expressing, Staining