anti egfr  (Abcam)

 
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    Anti EGFR antibody EP38Y
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    Structured Review

    Abcam anti egfr
    Proposed functional action of DUXAP 9‐206 in modulating NSCLC proliferation and metastasis. Lnc RNA DUXAP 9‐206 directly binds with <t>Cbl‐b</t> to augment <t>EGFR</t> signaling and promotes non‐small cell lung cancer progression

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    anti egfr - by Bioz Stars, 2021-05
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    1) Product Images from "Lnc RNA DUXAP9‐206 directly binds with Cbl‐b to augment EGFR signaling and promotes non‐small cell lung cancer progression, et al. LncRNA DUXAP9‐206 directly binds with Cbl‐b to augment EGFR signaling and promotes non‐small cell lung cancer progression"

    Article Title: Lnc RNA DUXAP9‐206 directly binds with Cbl‐b to augment EGFR signaling and promotes non‐small cell lung cancer progression, et al. LncRNA DUXAP9‐206 directly binds with Cbl‐b to augment EGFR signaling and promotes non‐small cell lung cancer progression

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.14085

    Proposed functional action of DUXAP 9‐206 in modulating NSCLC proliferation and metastasis. Lnc RNA DUXAP 9‐206 directly binds with Cbl‐b to augment EGFR signaling and promotes non‐small cell lung cancer progression
    Figure Legend Snippet: Proposed functional action of DUXAP 9‐206 in modulating NSCLC proliferation and metastasis. Lnc RNA DUXAP 9‐206 directly binds with Cbl‐b to augment EGFR signaling and promotes non‐small cell lung cancer progression

    Techniques Used: Functional Assay

    DUXAP 9‐206 interacts with Cbl‐b to reduce the degradation of EGFR . (A) The interaction between EGFR and Cbl‐b with DUXAP 9‐206 overexpression or knockdown was assessed by Co‐ IP assays. (B) Western blotting analysis of the expression levels of Cbl‐b and the EGFR signaling pathway downstream molecules p‐ AKT , AKT , p‐ ERK and ERK with ectopic expression or silencing of DUXAP 9‐206. Quantification of invading cells (C) and colony formation ability (D) in vector‐ and DUXAP 9‐206‐overexpressing cells by silencing of EGFR
    Figure Legend Snippet: DUXAP 9‐206 interacts with Cbl‐b to reduce the degradation of EGFR . (A) The interaction between EGFR and Cbl‐b with DUXAP 9‐206 overexpression or knockdown was assessed by Co‐ IP assays. (B) Western blotting analysis of the expression levels of Cbl‐b and the EGFR signaling pathway downstream molecules p‐ AKT , AKT , p‐ ERK and ERK with ectopic expression or silencing of DUXAP 9‐206. Quantification of invading cells (C) and colony formation ability (D) in vector‐ and DUXAP 9‐206‐overexpressing cells by silencing of EGFR

    Techniques Used: Over Expression, Co-Immunoprecipitation Assay, Western Blot, Expressing, Plasmid Preparation

    2) Product Images from "Destabilization of β-catenin and RAS by targeting the Wnt/β-catenin pathway as a potential treatment for triple-negative breast cancer"

    Article Title: Destabilization of β-catenin and RAS by targeting the Wnt/β-catenin pathway as a potential treatment for triple-negative breast cancer

    Journal: Experimental & Molecular Medicine

    doi: 10.1038/s12276-020-0440-y

    The increased expression and correlation of β-catenin, pan-RAS, and EGFR in the tumor tissues of TNBC patients. a IHC analyses of the TMA including TNBC tissues ( n = 34) and NATs ( n = 10) were performed using β-catenin, pan-RAS, or EGFR antibodies. Scale bar represents 200 μm. b Quantified H-score of nuclear β-catenin ( p
    Figure Legend Snippet: The increased expression and correlation of β-catenin, pan-RAS, and EGFR in the tumor tissues of TNBC patients. a IHC analyses of the TMA including TNBC tissues ( n = 34) and NATs ( n = 10) were performed using β-catenin, pan-RAS, or EGFR antibodies. Scale bar represents 200 μm. b Quantified H-score of nuclear β-catenin ( p

    Techniques Used: Expressing, Immunohistochemistry

    3) Product Images from "Linc00152 suppresses apoptosis and promotes migration by sponging miR-4767 in vascular endothelial cells"

    Article Title: Linc00152 suppresses apoptosis and promotes migration by sponging miR-4767 in vascular endothelial cells

    Journal: Oncotarget

    doi: 10.18632/oncotarget.18777

    MiR-4767 directly targets Bcl2L12 and EGFR (A) Potential sited targeted by miR-4767 in the sequences of Bcl2L12 and EGFR 3’UTRs. (B) wt and mut sequences of Bcl2L12 or EGFR inserted into the luciferase reporter gene vectors. Luc-Bcl2L12 3’UTR-wt or Luc-Bcl2L12 3’UTR-mut vectors (C) and Luc-EGFR 3’UTR-wt or Luc-EGFR 3’UTR-mut vectors (D) were co-transfected into HEK293T cells with 40 nM miR-4767 mimics or NC mimics for 48 h and luciferase activity was measured. ** P
    Figure Legend Snippet: MiR-4767 directly targets Bcl2L12 and EGFR (A) Potential sited targeted by miR-4767 in the sequences of Bcl2L12 and EGFR 3’UTRs. (B) wt and mut sequences of Bcl2L12 or EGFR inserted into the luciferase reporter gene vectors. Luc-Bcl2L12 3’UTR-wt or Luc-Bcl2L12 3’UTR-mut vectors (C) and Luc-EGFR 3’UTR-wt or Luc-EGFR 3’UTR-mut vectors (D) were co-transfected into HEK293T cells with 40 nM miR-4767 mimics or NC mimics for 48 h and luciferase activity was measured. ** P

    Techniques Used: Luciferase, Transfection, Activity Assay

    Linc00152 promoted the expression of Bcl2L12 and EGFR through diminishing miR-4767 (A) MiR-4767 negatively regulated the protein levels of Bcl2L12 and EGFR. HUVECs were transfected with miR-4767 mimics at concentrations of 20, and 40 nM or miR-4767 antagomirs at 10 and 20 nM for 48 h. Then, the cells were treated with 150 μg/mL ox-LDL for 24 h and the protein levels of Bcl2L12 and EGFR were detected by Western blotting. (B) Linc00152 promoted the expression of Bcl2L12 and EGFR through diminishing miR-4767. (C and D) Block of miR-4767 antagonized the changes of apoptosis and migration in HUVECs caused by linc00152 knockdown. HUVECs were transfected with 1.0 μg/mL pcDNA3.1-linc00152, 40 nM Linc00152 siRNA, or 40 nM Linc00152-siRNA plus 20 nM miR-4767 antagomirs. After incubation for 48 h, the cells were treated with 150 μg/mL ox-LDL for 24 h. The protein levels of Bcl2L12 and EGFR were detected by Western blotting. Cell apoptosis was checked by TUNEL assay and cell migration was detected with the Transwell Migration assay. * P
    Figure Legend Snippet: Linc00152 promoted the expression of Bcl2L12 and EGFR through diminishing miR-4767 (A) MiR-4767 negatively regulated the protein levels of Bcl2L12 and EGFR. HUVECs were transfected with miR-4767 mimics at concentrations of 20, and 40 nM or miR-4767 antagomirs at 10 and 20 nM for 48 h. Then, the cells were treated with 150 μg/mL ox-LDL for 24 h and the protein levels of Bcl2L12 and EGFR were detected by Western blotting. (B) Linc00152 promoted the expression of Bcl2L12 and EGFR through diminishing miR-4767. (C and D) Block of miR-4767 antagonized the changes of apoptosis and migration in HUVECs caused by linc00152 knockdown. HUVECs were transfected with 1.0 μg/mL pcDNA3.1-linc00152, 40 nM Linc00152 siRNA, or 40 nM Linc00152-siRNA plus 20 nM miR-4767 antagomirs. After incubation for 48 h, the cells were treated with 150 μg/mL ox-LDL for 24 h. The protein levels of Bcl2L12 and EGFR were detected by Western blotting. Cell apoptosis was checked by TUNEL assay and cell migration was detected with the Transwell Migration assay. * P

    Techniques Used: Expressing, Transfection, Western Blot, Blocking Assay, Migration, Incubation, TUNEL Assay, Transwell Migration Assay

    4) Product Images from "Dependence on the MUC1-C Oncoprotein in Non-Small Cell Lung Cancer Cells"

    Article Title: Dependence on the MUC1-C Oncoprotein in Non-Small Cell Lung Cancer Cells

    Journal: Molecular cancer therapeutics

    doi: 10.1158/1535-7163.MCT-10-1050

    EGFR mutant NSCLC cells are sensitive to treatment with MUC1-C inhibitors
    Figure Legend Snippet: EGFR mutant NSCLC cells are sensitive to treatment with MUC1-C inhibitors

    Techniques Used: Mutagenesis

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

    Article Title: Ectodysplasin A protein promotes corneal epithelial cell proliferation

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M117.803809

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

    Techniques Used: Cell Culture, CCK-8 Assay

    6) Product Images from "The MARCH Family E3 Ubiquitin Ligase K5 Alters Monocyte Metabolism and Proliferation through Receptor Tyrosine Kinase Modulation"

    Article Title: The MARCH Family E3 Ubiquitin Ligase K5 Alters Monocyte Metabolism and Proliferation through Receptor Tyrosine Kinase Modulation

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1001331

    K5 interacts with and alters RTK localization. Equal cell numbers of the indicated THP-1 lines were fixed with paraformaldehyde (PFA) and ( A ) stained without permeabilization to determine surface expression or ( B ) permeabilized with saponin prior to staining to determine total expression of Flt-4, PDGFR-ß, Flt-3 and EGFR by flow cytometry. Data are representative of three independent experiments. ( B, Inset ) The relative ratio of surface versus total RTKs was determined for vector- and K5 WT-expressing THP-1 cells. ( C ) 293T cells were co-transfected with expression constructs for EGFR, PDGFR-ß, or Flt-4 and the indicated GST expression constructs. After two days, lysates were subjected to GST pull-down using glutathione-sepharose beads. Purified proteins and whole cell lysates (WCL) were subjected to western blot (WB) using anti-EGFR, -Flt-4 or -PDGFR-ß antibodies, followed by re-probing with anti-GST antibodies. Arrows indicate GST or GST-K5 WT and mutant specific bands. Data are representative of three independent experiments.
    Figure Legend Snippet: K5 interacts with and alters RTK localization. Equal cell numbers of the indicated THP-1 lines were fixed with paraformaldehyde (PFA) and ( A ) stained without permeabilization to determine surface expression or ( B ) permeabilized with saponin prior to staining to determine total expression of Flt-4, PDGFR-ß, Flt-3 and EGFR by flow cytometry. Data are representative of three independent experiments. ( B, Inset ) The relative ratio of surface versus total RTKs was determined for vector- and K5 WT-expressing THP-1 cells. ( C ) 293T cells were co-transfected with expression constructs for EGFR, PDGFR-ß, or Flt-4 and the indicated GST expression constructs. After two days, lysates were subjected to GST pull-down using glutathione-sepharose beads. Purified proteins and whole cell lysates (WCL) were subjected to western blot (WB) using anti-EGFR, -Flt-4 or -PDGFR-ß antibodies, followed by re-probing with anti-GST antibodies. Arrows indicate GST or GST-K5 WT and mutant specific bands. Data are representative of three independent experiments.

    Techniques Used: Staining, Expressing, Flow Cytometry, Cytometry, Plasmid Preparation, Transfection, Construct, Purification, Western Blot, Mutagenesis

    7) Product Images from "Exosome-delivered EGFR regulates liver microenvironment to promote gastric cancer liver metastasis"

    Article Title: Exosome-delivered EGFR regulates liver microenvironment to promote gastric cancer liver metastasis

    Journal: Nature Communications

    doi: 10.1038/ncomms15016

    SGC-exosomes-mediated EGFR activates liver HGF by suppressing miR-26a/b expression. Forty micrograms of exosomes were used to culture with 1 × 10 6 primary liver cells seeded in a six-well plate. ( a , b ) Effects of SGC exosomes delivered EGFR on HGF protein levels ( a ) and mRNA levels ( b ) in mixed primary liver cells ( n =3). ( c , d ) Effects of EGFR on the expression of HGF protein ( c ) and mRNA ( d ) ( n =3). ( e ) The binding sites of miR-26a/b in the 3′-UTR of HGF mRNA. ( f ) SGC-exosomes decrease liver miR-26a/b levels ( n =3). ( g ) EGFR is negatively related with miR-26a/b in liver cells ( n =3). ( h ) Absolute quantification of serum miR-26a/b in the progression of GC ( n =150). The data represent the mean±s.e.m. ** P
    Figure Legend Snippet: SGC-exosomes-mediated EGFR activates liver HGF by suppressing miR-26a/b expression. Forty micrograms of exosomes were used to culture with 1 × 10 6 primary liver cells seeded in a six-well plate. ( a , b ) Effects of SGC exosomes delivered EGFR on HGF protein levels ( a ) and mRNA levels ( b ) in mixed primary liver cells ( n =3). ( c , d ) Effects of EGFR on the expression of HGF protein ( c ) and mRNA ( d ) ( n =3). ( e ) The binding sites of miR-26a/b in the 3′-UTR of HGF mRNA. ( f ) SGC-exosomes decrease liver miR-26a/b levels ( n =3). ( g ) EGFR is negatively related with miR-26a/b in liver cells ( n =3). ( h ) Absolute quantification of serum miR-26a/b in the progression of GC ( n =150). The data represent the mean±s.e.m. ** P

    Techniques Used: Expressing, Binding Assay

    Clinical analysis of sr-exosome EGFR and HGF-cMET in GC. ( a ) Electron microscope scanning of exosomes isolated from human serum. ( b ) Sr-exosome EGFR is related to the progression of GC. Exosomes were isolated from serum of healthy donors (NC), stage II/III GC patients and stage IV GC patients, respectively ( n =20); Alix, TSG101 and CD63 were used as the internal control of exosomes. ( c ) The expression of HGF, c-MET and p-c-MET in para-carcinoma tissue (liver) and GC liver metastases ( n =5). ( d ) Immunohistochemistry (IHC) analysis of HGF in GC liver metastasis. ( e ) Positive ratio of HGF and its receptor c-MET in GC liver metastases ( n =30). The data represent the mean±s.e.m. * P
    Figure Legend Snippet: Clinical analysis of sr-exosome EGFR and HGF-cMET in GC. ( a ) Electron microscope scanning of exosomes isolated from human serum. ( b ) Sr-exosome EGFR is related to the progression of GC. Exosomes were isolated from serum of healthy donors (NC), stage II/III GC patients and stage IV GC patients, respectively ( n =20); Alix, TSG101 and CD63 were used as the internal control of exosomes. ( c ) The expression of HGF, c-MET and p-c-MET in para-carcinoma tissue (liver) and GC liver metastases ( n =5). ( d ) Immunohistochemistry (IHC) analysis of HGF in GC liver metastasis. ( e ) Positive ratio of HGF and its receptor c-MET in GC liver metastases ( n =30). The data represent the mean±s.e.m. * P

    Techniques Used: Microscopy, Isolation, Expressing, Immunohistochemistry

    In vivo verification for exosome-EGFR activates liver HGF by inhibiting miR-26a/b. The livers of mice were pre-treated with HGF shRNA or HGF overexpressing lentivirus by multi-point injection, followed by orthotopic tumour implantation on the 6th day; finally, mice were killed and data were analysed on the 66th day. ( a ) A flow chart depicting the in vivo experimental design. ( b ) Immunohistochemistry (IHC) analysis of EGFR expression in primary gastric tumour. ( c ) Western blotting analysis of exosome EGFR in the serum of tumour-implanted mice ( n =30). ( d ) Levels of HGF in liver tissues of GC tumour-implanted mice ( n =30). ( e ) Electron-microscope scanning of exosomes isolated from mouse serum. ( f ) Relative levels of miR-26a/b in mouse liver tissues ( n =30). The data represent the mean±s.e.m. ** P
    Figure Legend Snippet: In vivo verification for exosome-EGFR activates liver HGF by inhibiting miR-26a/b. The livers of mice were pre-treated with HGF shRNA or HGF overexpressing lentivirus by multi-point injection, followed by orthotopic tumour implantation on the 6th day; finally, mice were killed and data were analysed on the 66th day. ( a ) A flow chart depicting the in vivo experimental design. ( b ) Immunohistochemistry (IHC) analysis of EGFR expression in primary gastric tumour. ( c ) Western blotting analysis of exosome EGFR in the serum of tumour-implanted mice ( n =30). ( d ) Levels of HGF in liver tissues of GC tumour-implanted mice ( n =30). ( e ) Electron-microscope scanning of exosomes isolated from mouse serum. ( f ) Relative levels of miR-26a/b in mouse liver tissues ( n =30). The data represent the mean±s.e.m. ** P

    Techniques Used: In Vivo, Mouse Assay, shRNA, Injection, Flow Cytometry, Immunohistochemistry, Expressing, Western Blot, Microscopy, Isolation

    8) Product Images from "Combination of curcumin and ginkgolide B inhibits cystogenesis by regulating multiple signaling pathways"

    Article Title: Combination of curcumin and ginkgolide B inhibits cystogenesis by regulating multiple signaling pathways

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2021.11834

    Cur combined with GB regulates the EGFR/ERK1/2 signaling pathway in Pkd1 Flox/− ; Ksp-Cre mice. (A) Representative immunohistochemistry of p-EGFR and p-Cerb-B2 in Pkd1 Flox/− ; Ksp-Cre mice treated with 160 mg/kg Cur, 80 mg/kg GB and 160 mg/kg Cur combined with 80 mg/kg GB. Magnification, ×400. (B) Representative western blotting of EGFR/ERK1/2 signaling proteins in Pkd1 Flox/− ; Ksp-Cre mice treated with 160 mg/kg Cur, 80 mg/kg GB and 160 mg/kg Cur combined with 80 mg/kg GB. (C) Semi-quantitative analysis of EGFR/ERK1/2 signaling protein expression levels in Pkd1 Flox/− ; Ksp-Cre mice. Relative level refers to the ratio of western blotting band density in different treatment groups compared with that in WT group. Data are presented as the mean ± SEM, n=6. *P
    Figure Legend Snippet: Cur combined with GB regulates the EGFR/ERK1/2 signaling pathway in Pkd1 Flox/− ; Ksp-Cre mice. (A) Representative immunohistochemistry of p-EGFR and p-Cerb-B2 in Pkd1 Flox/− ; Ksp-Cre mice treated with 160 mg/kg Cur, 80 mg/kg GB and 160 mg/kg Cur combined with 80 mg/kg GB. Magnification, ×400. (B) Representative western blotting of EGFR/ERK1/2 signaling proteins in Pkd1 Flox/− ; Ksp-Cre mice treated with 160 mg/kg Cur, 80 mg/kg GB and 160 mg/kg Cur combined with 80 mg/kg GB. (C) Semi-quantitative analysis of EGFR/ERK1/2 signaling protein expression levels in Pkd1 Flox/− ; Ksp-Cre mice. Relative level refers to the ratio of western blotting band density in different treatment groups compared with that in WT group. Data are presented as the mean ± SEM, n=6. *P

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

    9) Product Images from "Cells with intense EGFR staining and a high nuclear to cytoplasmic ratio are specific for infiltrative glioma: a useful marker in neuropathological practice"

    Article Title: Cells with intense EGFR staining and a high nuclear to cytoplasmic ratio are specific for infiltrative glioma: a useful marker in neuropathological practice

    Journal: Neuro-Oncology

    doi: 10.1093/neuonc/not094

    (A) Characterization of strongly EGFR-positive cells in infiltrative glioma tissue sections. Double-staining was performed on paraffin-embedded tissue section of infiltrative glioma using anti-EGFR antibody (green), and (a) anti-GFAP antibody (red) or (b) anti-olig2 antibody (red), or (c) anti-nestin antibody (red) (×400). Strongly EGFR-stained cells with a high nucleus to cytoplasm ratio lacked GFAP expression (a) and coexpressed olig2 (b) and nestin (c). (B and C) Characterization of EGFR-overexpressing cells sorted from fresh glioblastoma. Immunofluorescence performed immediately after FACS sorting of small EGFR-overexpressing cells (B) confirmed that cells overexpressed EGFR and showed that these cells expressed the neural stem cell markers oct4, sox1, sox2, and A2B5 and (C) expressed invasion marker CXCR4 and CD44 (×1000). (D) Culture of small EGFR overexpressing cells in a medium specific for neural stem cells (×1000). Cells slowly proliferated and formed small neurospheres (left) or remained nonadhesive quiescent isolated cells (right).
    Figure Legend Snippet: (A) Characterization of strongly EGFR-positive cells in infiltrative glioma tissue sections. Double-staining was performed on paraffin-embedded tissue section of infiltrative glioma using anti-EGFR antibody (green), and (a) anti-GFAP antibody (red) or (b) anti-olig2 antibody (red), or (c) anti-nestin antibody (red) (×400). Strongly EGFR-stained cells with a high nucleus to cytoplasm ratio lacked GFAP expression (a) and coexpressed olig2 (b) and nestin (c). (B and C) Characterization of EGFR-overexpressing cells sorted from fresh glioblastoma. Immunofluorescence performed immediately after FACS sorting of small EGFR-overexpressing cells (B) confirmed that cells overexpressed EGFR and showed that these cells expressed the neural stem cell markers oct4, sox1, sox2, and A2B5 and (C) expressed invasion marker CXCR4 and CD44 (×1000). (D) Culture of small EGFR overexpressing cells in a medium specific for neural stem cells (×1000). Cells slowly proliferated and formed small neurospheres (left) or remained nonadhesive quiescent isolated cells (right).

    Techniques Used: Double Staining, Staining, Expressing, Immunofluorescence, FACS, Marker, Isolation

    10) Product Images from "A novel co-drug of aspirin and ursolic acid interrupts adhesion, invasion and migration of cancer cells to vascular endothelium via regulating EMT and EGFR-mediated signaling pathways: multiple targets for cancer metastasis prevention and treatment"

    Article Title: A novel co-drug of aspirin and ursolic acid interrupts adhesion, invasion and migration of cancer cells to vascular endothelium via regulating EMT and EGFR-mediated signaling pathways: multiple targets for cancer metastasis prevention and treatment

    Journal: Oncotarget

    doi: 10.18632/oncotarget.12232

    Influence of Asp-UA on the cell invasion molecules, EMT and EGFR related cell signaling pathways ( A ) Western blot analysis of UA/Asp/Asp-UA on the expression of MMP-2, MMP-9 and COX-2 in MCF-7 cells and ( B ) MDA-MB-231 cells. ( C ) Expression of E-cadherin on MCF-7 cells was determined by flow cytometry, isotype control (gray area), control (black curve), blue curve (15 μM UA), yellow curve (1 mM Asp) and red line represents Asp-UA treated group at concentrations of 20 and 40 μM, respectively. ( D ) Western blotting analysis of UA/Asp/Asp-UA on the regulation of E-cadherin, β-catenin, EGFR, ERK and PTEN in MCF-7 ( E ) and MDA-MB-231 cells ( F ). The inhibitory effects of UA, Asp and Asp-UA on the expression of E-cadherin by FACS. ( G ) Double immunofluorescence staining with DAPI (blue) and anti-EGFR antibody (red) was carried out in MCF-7 cells treated with Asp-UA for 24 h. The data were obtained from 3 separate experiments and bars represent the mean ± SD. (* P
    Figure Legend Snippet: Influence of Asp-UA on the cell invasion molecules, EMT and EGFR related cell signaling pathways ( A ) Western blot analysis of UA/Asp/Asp-UA on the expression of MMP-2, MMP-9 and COX-2 in MCF-7 cells and ( B ) MDA-MB-231 cells. ( C ) Expression of E-cadherin on MCF-7 cells was determined by flow cytometry, isotype control (gray area), control (black curve), blue curve (15 μM UA), yellow curve (1 mM Asp) and red line represents Asp-UA treated group at concentrations of 20 and 40 μM, respectively. ( D ) Western blotting analysis of UA/Asp/Asp-UA on the regulation of E-cadherin, β-catenin, EGFR, ERK and PTEN in MCF-7 ( E ) and MDA-MB-231 cells ( F ). The inhibitory effects of UA, Asp and Asp-UA on the expression of E-cadherin by FACS. ( G ) Double immunofluorescence staining with DAPI (blue) and anti-EGFR antibody (red) was carried out in MCF-7 cells treated with Asp-UA for 24 h. The data were obtained from 3 separate experiments and bars represent the mean ± SD. (* P

    Techniques Used: Western Blot, Expressing, Multiple Displacement Amplification, Flow Cytometry, Cytometry, FACS, Double Immunofluorescence Staining

    11) Product Images from "Epidermal Growth Factor Receptor (EGFR) Signaling Regulates Epiphyseal Cartilage Development through β-Catenin-dependent and -independent Pathways *"

    Article Title: Epidermal Growth Factor Receptor (EGFR) Signaling Regulates Epiphyseal Cartilage Development through β-Catenin-dependent and -independent Pathways *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M113.463554

    The development of SOC in the proximal epiphysis of the tibia is delayed in the chondrocyte-specific EGFR-deficient mice. A , immunohistochemistry of EGFR in the epiphyseal cartilage of P9 WT mice reveals that most chondrocytes express EGFR ( a ). Black
    Figure Legend Snippet: The development of SOC in the proximal epiphysis of the tibia is delayed in the chondrocyte-specific EGFR-deficient mice. A , immunohistochemistry of EGFR in the epiphyseal cartilage of P9 WT mice reveals that most chondrocytes express EGFR ( a ). Black

    Techniques Used: Mouse Assay, Immunohistochemistry

    EGFR signaling stimulates the expression of Mmp9 and -13 and Rankl through β-catenin-dependent and -independent pathways. A , TGFα stimulates the expression of Mmp9 and -13 and Rankl in mouse chondrocytes through the EGFR pathway. Mouse
    Figure Legend Snippet: EGFR signaling stimulates the expression of Mmp9 and -13 and Rankl through β-catenin-dependent and -independent pathways. A , TGFα stimulates the expression of Mmp9 and -13 and Rankl in mouse chondrocytes through the EGFR pathway. Mouse

    Techniques Used: Expressing

    EGFR deficiency in chondrocytes suppresses the terminal differentiation and apoptosis of hypertrophic chondrocytes adjacent to the marrow space in the SOC. A , immunohistochemistry of Runx2 protein ( a–c ) and TUNEL ( d–f ) staining in sections
    Figure Legend Snippet: EGFR deficiency in chondrocytes suppresses the terminal differentiation and apoptosis of hypertrophic chondrocytes adjacent to the marrow space in the SOC. A , immunohistochemistry of Runx2 protein ( a–c ) and TUNEL ( d–f ) staining in sections

    Techniques Used: Immunohistochemistry, TUNEL Assay, Staining

    Blocking EGFR activity in chondrocytes reduces the amounts of MMPs and cartilage matrix degradation in the hypertrophic chondrocytes adjacent to the marrow space in the SOC. A , immunohistochemistry of MMP9 ( a–c ), 13 ( d–f ), 14 ( g–i
    Figure Legend Snippet: Blocking EGFR activity in chondrocytes reduces the amounts of MMPs and cartilage matrix degradation in the hypertrophic chondrocytes adjacent to the marrow space in the SOC. A , immunohistochemistry of MMP9 ( a–c ), 13 ( d–f ), 14 ( g–i

    Techniques Used: Blocking Assay, Activity Assay, Immunohistochemistry

    12) Product Images from "miR-2861 acts as a tumor suppressor via targeting EGFR/AKT2/CCND1 pathway in cervical cancer induced by human papillomavirus virus 16 E6"

    Article Title: miR-2861 acts as a tumor suppressor via targeting EGFR/AKT2/CCND1 pathway in cervical cancer induced by human papillomavirus virus 16 E6

    Journal: Scientific Reports

    doi: 10.1038/srep28968

    Knockdown of EGFR, AKT2, or CCND1 produces similar suppressive effects to that of miR-2861 overexpression in cervical cancer cells. ( A ) Knockdown of EGFR, AKT2, or CCND1 suppresses cell proliferation of both SiHa and CaSki cells. CCK8 assay was performed to determine the growth of SiHa and CaSki cells treated with si-EGFR, si-AKT2, si-CCND1, or si-NS. ( B , C ) Knockdown of EGFR, AKT2, or CCND1 enhances cell apoptosis of SiHa and CaSki cells. Apoptosis assay was determined in SiHa and CaSki cells at 48 h after transfection of si-EGFR, si-AKT2, si-CCND1, or si-NS. Representative images are shown ( C ) and early apoptosis rate are indicated ( B ). ** P
    Figure Legend Snippet: Knockdown of EGFR, AKT2, or CCND1 produces similar suppressive effects to that of miR-2861 overexpression in cervical cancer cells. ( A ) Knockdown of EGFR, AKT2, or CCND1 suppresses cell proliferation of both SiHa and CaSki cells. CCK8 assay was performed to determine the growth of SiHa and CaSki cells treated with si-EGFR, si-AKT2, si-CCND1, or si-NS. ( B , C ) Knockdown of EGFR, AKT2, or CCND1 enhances cell apoptosis of SiHa and CaSki cells. Apoptosis assay was determined in SiHa and CaSki cells at 48 h after transfection of si-EGFR, si-AKT2, si-CCND1, or si-NS. Representative images are shown ( C ) and early apoptosis rate are indicated ( B ). ** P

    Techniques Used: Over Expression, CCK-8 Assay, Apoptosis Assay, Transfection

    miR-2861 targets EGFR, AKT2, and CCND1 in cervical cancer cells. ( A ) Predicted EGFR, AKT2, and CCND1 3′ UTR binding sites for miR-2861. The alignment of the seed region of miR-2861 with EGFR, AKT2, and CCND1 3′UTR are shown. The mutated sites of targets are indicated in red. ( B ) EGFR 3′UTR is a target of miR-2861. pmiR-GLO luciferase construct containing a wt (left) or mutated (right) EGFR 3′UTR was cotransfected with miR-2861 or miR-NC in SiHa cells and the luciferase reporter assay was performed at 24 h posttransfection. ( C ) AKT2 3′UTR is a target of miR-2861. pmiR-GLO luciferase construct containing each wt (wt1, 2, 3, 4, and 5) or mutated (mut1, 2, 3, 4, and 5) AKT2 3′UTR was cotransfected with miR-2861 or miR-NC in SiHa cells, respectively. The luciferase assay was then performed. ( D ) CCND1 3′UTR is also a target of miR-2861. pmiR-GLO luciferase construct containing a wt (left) or mutated (right) CCND1 3′UTR was cotransfected with miR-2861 or miR-NC and the luciferase assay was performed. ( E ) miR-2861 overexpression decreased endogenous levels of EGFR, AKT2, and CCND1 proteins in SiHa and CaSki cells. SiHa and CaSki cells were transfected with miR-2861 or miR-NC for 72 h, respectively. EGFR, AKT2, and CCND1 expressions were assessed by Western blot. GAPDH was obtained as a loading control. Error bars, ± SD. * P
    Figure Legend Snippet: miR-2861 targets EGFR, AKT2, and CCND1 in cervical cancer cells. ( A ) Predicted EGFR, AKT2, and CCND1 3′ UTR binding sites for miR-2861. The alignment of the seed region of miR-2861 with EGFR, AKT2, and CCND1 3′UTR are shown. The mutated sites of targets are indicated in red. ( B ) EGFR 3′UTR is a target of miR-2861. pmiR-GLO luciferase construct containing a wt (left) or mutated (right) EGFR 3′UTR was cotransfected with miR-2861 or miR-NC in SiHa cells and the luciferase reporter assay was performed at 24 h posttransfection. ( C ) AKT2 3′UTR is a target of miR-2861. pmiR-GLO luciferase construct containing each wt (wt1, 2, 3, 4, and 5) or mutated (mut1, 2, 3, 4, and 5) AKT2 3′UTR was cotransfected with miR-2861 or miR-NC in SiHa cells, respectively. The luciferase assay was then performed. ( D ) CCND1 3′UTR is also a target of miR-2861. pmiR-GLO luciferase construct containing a wt (left) or mutated (right) CCND1 3′UTR was cotransfected with miR-2861 or miR-NC and the luciferase assay was performed. ( E ) miR-2861 overexpression decreased endogenous levels of EGFR, AKT2, and CCND1 proteins in SiHa and CaSki cells. SiHa and CaSki cells were transfected with miR-2861 or miR-NC for 72 h, respectively. EGFR, AKT2, and CCND1 expressions were assessed by Western blot. GAPDH was obtained as a loading control. Error bars, ± SD. * P

    Techniques Used: Binding Assay, Luciferase, Construct, Reporter Assay, Over Expression, Transfection, Western Blot

    Knockdown of EGFR, AKT2, or CCND1 suppresses cell invasion and increased E-cadherin level in cervical cancer cells. ( A , B ) A matrigel invasion assay was performed in SiHa ( A ) and CaSki ( B ) cells transfected with si-EGFR, si-AKT2, si-CCND1, or si-NS. Representative images are shown (magnification: ×200). Error bars indicate ± SD. ( C ) Suppression of EGFR, AKT2, or CCND1 increased E-cadherin expression level in both SiHa and CaSki cells. 72 h after transfection, the protein level of E-cadherin was analyzed by Western blot. ** P
    Figure Legend Snippet: Knockdown of EGFR, AKT2, or CCND1 suppresses cell invasion and increased E-cadherin level in cervical cancer cells. ( A , B ) A matrigel invasion assay was performed in SiHa ( A ) and CaSki ( B ) cells transfected with si-EGFR, si-AKT2, si-CCND1, or si-NS. Representative images are shown (magnification: ×200). Error bars indicate ± SD. ( C ) Suppression of EGFR, AKT2, or CCND1 increased E-cadherin expression level in both SiHa and CaSki cells. 72 h after transfection, the protein level of E-cadherin was analyzed by Western blot. ** P

    Techniques Used: Invasion Assay, Transfection, Expressing, Western Blot

    EGFR, AKT2, and CCND1 rescue miR-2861-induced cellular phenotypes in SiHa cells. Cells were cotransfected with EGFR, AKT2, CCND1 or empty vector and miR-2861 or miR-NC. ( A ) Overexpression of either EGFR, AKT2, or CCND1 recovers miR-2861-induced cell proliferation. CCK8 assay was assessed at 72 h after cotransfection. ( B ) Overexpression of either EGFR, AKT2, or CCND1 rescues miR-2861-induced inhibition of cell invasion. Representative images are shown (magnification: ×200). Error bars indicate ± SD. * P
    Figure Legend Snippet: EGFR, AKT2, and CCND1 rescue miR-2861-induced cellular phenotypes in SiHa cells. Cells were cotransfected with EGFR, AKT2, CCND1 or empty vector and miR-2861 or miR-NC. ( A ) Overexpression of either EGFR, AKT2, or CCND1 recovers miR-2861-induced cell proliferation. CCK8 assay was assessed at 72 h after cotransfection. ( B ) Overexpression of either EGFR, AKT2, or CCND1 rescues miR-2861-induced inhibition of cell invasion. Representative images are shown (magnification: ×200). Error bars indicate ± SD. * P

    Techniques Used: Plasmid Preparation, Over Expression, CCK-8 Assay, Cotransfection, Inhibition

    13) Product Images from "The role of c-Src in the invasion and metastasis of hepatocellular carcinoma cells induced by association of cell surface GRP78 with activated α2M"

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

    Journal: BMC Cancer

    doi: 10.1186/s12885-015-1401-z

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

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

    14) Product Images from "Phosphoproteomic analysis reveals Smarcb1 dependent EGFR signaling in Malignant Rhabdoid tumor cells"

    Article Title: Phosphoproteomic analysis reveals Smarcb1 dependent EGFR signaling in Malignant Rhabdoid tumor cells

    Journal: Molecular Cancer

    doi: 10.1186/s12943-015-0439-5

    EGFR activation mediates AKT activation in Smarcb1 deficient cells. a Inhibition of AKT activation upon treatment with the EGFR/HER2 inhibitor Lapatinib (Lap.) and the EGFR inhibitor Gefitinib (Gef.) versus the DMSO (D.) control. b WST1 proliferation assay demonstrating relative proliferation of Smarcb1 deficient and proficient cells following a 6 day treatment with the EGFR inhibitors Lapatinib/Gefitinib, with the AKT inhibitor 1/2, dual inhibition and serum withdrawal
    Figure Legend Snippet: EGFR activation mediates AKT activation in Smarcb1 deficient cells. a Inhibition of AKT activation upon treatment with the EGFR/HER2 inhibitor Lapatinib (Lap.) and the EGFR inhibitor Gefitinib (Gef.) versus the DMSO (D.) control. b WST1 proliferation assay demonstrating relative proliferation of Smarcb1 deficient and proficient cells following a 6 day treatment with the EGFR inhibitors Lapatinib/Gefitinib, with the AKT inhibitor 1/2, dual inhibition and serum withdrawal

    Techniques Used: Activation Assay, Inhibition, Proliferation Assay

    15) Product Images from "Rhythmic Pressure Waves Induce Mucin5AC Expression via an EGFR-Mediated Signaling Pathway in Human Airway Epithelial Cells"

    Article Title: Rhythmic Pressure Waves Induce Mucin5AC Expression via an EGFR-Mediated Signaling Pathway in Human Airway Epithelial Cells

    Journal: DNA and Cell Biology

    doi: 10.1089/dna.2013.2079

    The effect of RPW on the activation of EGFR, ERK1/2, and Akt in 16HBE cells. Cell lysates were prepared after 2 h of RPW exposure; p-EGFR (A) , p-ERK1/2 (B) , and p-Akt (C) protein expressions were measured by Western blotting analyses using specific
    Figure Legend Snippet: The effect of RPW on the activation of EGFR, ERK1/2, and Akt in 16HBE cells. Cell lysates were prepared after 2 h of RPW exposure; p-EGFR (A) , p-ERK1/2 (B) , and p-Akt (C) protein expressions were measured by Western blotting analyses using specific

    Techniques Used: Activation Assay, Western Blot

    The role of EGFR in RPW induced the phosphorylation of ERK1/2 and Akt. The cells were transfected with a siNT or specific siRNA for EGFR (siEGFR) before applying RPW. Cell lysates were prepared after 2 h of RPW exposure; p-ERK1/2 (A) and p-Akt
    Figure Legend Snippet: The role of EGFR in RPW induced the phosphorylation of ERK1/2 and Akt. The cells were transfected with a siNT or specific siRNA for EGFR (siEGFR) before applying RPW. Cell lysates were prepared after 2 h of RPW exposure; p-ERK1/2 (A) and p-Akt

    Techniques Used: Transfection

    16) Product Images from "The Effects of Artesunate on the Expression of EGFR and ABCG2 in A549 Human Lung Cancer Cells and a Xenograft Model"

    Article Title: The Effects of Artesunate on the Expression of EGFR and ABCG2 in A549 Human Lung Cancer Cells and a Xenograft Model

    Journal: Molecules

    doi: 10.3390/molecules161210556

    The expression of ABCG2, Akt, p-Akt and p-EGFR proteins was examined in untreated and ART-treated (60 mg·kg −1 and 120 mg·kg −1 ) xenografts. Photographs of immunohistochemical staining for ABCG2, Akt, p-Akt and p-EGFR (×40) are shown. A distinct yellow stain can be observed in the membrane and cytoplasm. The images presented are representative of three independent experiments. The data are the means ± SE from three independent experiments performed in triplicate. * p
    Figure Legend Snippet: The expression of ABCG2, Akt, p-Akt and p-EGFR proteins was examined in untreated and ART-treated (60 mg·kg −1 and 120 mg·kg −1 ) xenografts. Photographs of immunohistochemical staining for ABCG2, Akt, p-Akt and p-EGFR (×40) are shown. A distinct yellow stain can be observed in the membrane and cytoplasm. The images presented are representative of three independent experiments. The data are the means ± SE from three independent experiments performed in triplicate. * p

    Techniques Used: Expressing, Immunohistochemistry, Staining

    ART down-regulates the EGFR and ABCG2 mRNA and protein levels in A549 cells. ( A ) Cells were treated with varying concentrations of ART for 48 h and then harvested. The EGFR and ABCG2 mRNA levels were detected using real-time RT-PCR; ( B ) The expression of EGFR, p-EGFR Akt, p-Akt and ABCG2 was measured by western blot analysis. A549 cells were treated with varying concentrations of ART for 48 h, then β-actin expression was used as an internal control to determine the expression levels of proteins. Significant down-regulation of all proteins was observed. The blots are representative of three independent experiments. The data represent the means ± SE from three independent experiments performed in triplicate. * p
    Figure Legend Snippet: ART down-regulates the EGFR and ABCG2 mRNA and protein levels in A549 cells. ( A ) Cells were treated with varying concentrations of ART for 48 h and then harvested. The EGFR and ABCG2 mRNA levels were detected using real-time RT-PCR; ( B ) The expression of EGFR, p-EGFR Akt, p-Akt and ABCG2 was measured by western blot analysis. A549 cells were treated with varying concentrations of ART for 48 h, then β-actin expression was used as an internal control to determine the expression levels of proteins. Significant down-regulation of all proteins was observed. The blots are representative of three independent experiments. The data represent the means ± SE from three independent experiments performed in triplicate. * p

    Techniques Used: Quantitative RT-PCR, Expressing, Western Blot

    17) Product Images from "CD60b: Enriching Neural Stem/Progenitor Cells from Rat Development into Adulthood"

    Article Title: CD60b: Enriching Neural Stem/Progenitor Cells from Rat Development into Adulthood

    Journal: Stem Cells International

    doi: 10.1155/2017/5759490

    CD60b is expressed in different populations in the adult SVZ. Confocal images of adult SVZ sections showing the expression of CD60b (a, d, g, j, and m in red) and doublecortin (b-c), EGFR (e-f), GFAP (h-i), PNA (k-l), and vimentin (n-o) (in green). Nuclei were counterstained with TO-PRO-3 (blue). We observed the expression of CD60b in DCX (expressed in neuroblasts), R-EGF (expressed mostly in type C cells), GFAP (expressed in type B cells and astrocytes), and vimentin-positive cells (expressed in radial glia-like cells) (arrows). PNA, which was described as a negative marker of neural stem cells, and CD60b showed little colocalization (l). High magnification is showed in the insets. LV: lateral ventricle; DCX: doublecortin. Scale bar: 20 μ m.
    Figure Legend Snippet: CD60b is expressed in different populations in the adult SVZ. Confocal images of adult SVZ sections showing the expression of CD60b (a, d, g, j, and m in red) and doublecortin (b-c), EGFR (e-f), GFAP (h-i), PNA (k-l), and vimentin (n-o) (in green). Nuclei were counterstained with TO-PRO-3 (blue). We observed the expression of CD60b in DCX (expressed in neuroblasts), R-EGF (expressed mostly in type C cells), GFAP (expressed in type B cells and astrocytes), and vimentin-positive cells (expressed in radial glia-like cells) (arrows). PNA, which was described as a negative marker of neural stem cells, and CD60b showed little colocalization (l). High magnification is showed in the insets. LV: lateral ventricle; DCX: doublecortin. Scale bar: 20 μ m.

    Techniques Used: Expressing, Marker

    18) Product Images from "EGFR‐rich extracellular vesicles derived from highly metastatic nasopharyngeal carcinoma cells accelerate tumour metastasis through PI3K/AKT pathway‐suppressed ROS, et al. EGFR‐rich extracellular vesicles derived from highly metastatic nasopharyngeal carcinoma cells accelerate tumour metastasis through PI3K/AKT pathway‐suppressed ROS"

    Article Title: EGFR‐rich extracellular vesicles derived from highly metastatic nasopharyngeal carcinoma cells accelerate tumour metastasis through PI3K/AKT pathway‐suppressed ROS, et al. EGFR‐rich extracellular vesicles derived from highly metastatic nasopharyngeal carcinoma cells accelerate tumour metastasis through PI3K/AKT pathway‐suppressed ROS

    Journal: Journal of Extracellular Vesicles

    doi: 10.1002/jev2.12003

    EVs transfer from highly metastatic NPC cells to poorly metastatic NPC cells mediates cell‐cell communication. (a) Schematic representation of highly and poorly metastatic NPC cells co‐culture system. pmCherry‐CD63‐transfected highly metastatic NPC cells were seeded into the upper chamber and poorly metastatic NPC cells were seeded into the lower chamber. (b) Confocal microscopy observation of the internalization of EVs secreted from mCherry‐CD63‐labeled 5–8F or S18 (upper chamber) in 6–10B or S26 cells (lower chamber). Hoechst 33342 was used to stain cellular nuclei (blue fluorescence). Stereoscopic structure of cells was observed by differential interference contrast (DIC) (Magnification × 630; Scale bar, 10 μm). (c) Western blot analysis showing the presence of CD9, CD63, ALIX, TSG101, and EGFR and absence of GAPDH in EVs derived from the conditioned medium of NPC cells. (d) Representative nanoparticle tracking analysis plots showing the size distribution and total number of EVs isolated from the same volume of conditioned medium of 5–8F and S18 cells. (e) Confocal microscopy image showing the internalization of PKH26‐labeled EVs (red) isolated from the conditioned medium of highly metastatic NPC cells by 6–10B and S26 cells. Hoechst 33342 was used to stain the nuclei of cells (blue). Cell membrane was stained by Dio (green fluorescence). (Magnification × 630; Scale bar, 10 μm). Experiments were performed in triplicate
    Figure Legend Snippet: EVs transfer from highly metastatic NPC cells to poorly metastatic NPC cells mediates cell‐cell communication. (a) Schematic representation of highly and poorly metastatic NPC cells co‐culture system. pmCherry‐CD63‐transfected highly metastatic NPC cells were seeded into the upper chamber and poorly metastatic NPC cells were seeded into the lower chamber. (b) Confocal microscopy observation of the internalization of EVs secreted from mCherry‐CD63‐labeled 5–8F or S18 (upper chamber) in 6–10B or S26 cells (lower chamber). Hoechst 33342 was used to stain cellular nuclei (blue fluorescence). Stereoscopic structure of cells was observed by differential interference contrast (DIC) (Magnification × 630; Scale bar, 10 μm). (c) Western blot analysis showing the presence of CD9, CD63, ALIX, TSG101, and EGFR and absence of GAPDH in EVs derived from the conditioned medium of NPC cells. (d) Representative nanoparticle tracking analysis plots showing the size distribution and total number of EVs isolated from the same volume of conditioned medium of 5–8F and S18 cells. (e) Confocal microscopy image showing the internalization of PKH26‐labeled EVs (red) isolated from the conditioned medium of highly metastatic NPC cells by 6–10B and S26 cells. Hoechst 33342 was used to stain the nuclei of cells (blue). Cell membrane was stained by Dio (green fluorescence). (Magnification × 630; Scale bar, 10 μm). Experiments were performed in triplicate

    Techniques Used: Co-Culture Assay, Transfection, Confocal Microscopy, Labeling, Staining, Fluorescence, Western Blot, Derivative Assay, Isolation

    H‐EVs‐induced EGFR expression down‐regulates ROS via the PI3K/AKT pathway to promote metastasis of NPC cells. (a) Western blot analysis showing the presence of CD9, CD63, ALIX, and TSG101 and absence of GAPDH in EGFR‐KO EVs derived from the conditioned medium of EGFR‐KO 5–8F and S18 cells. (b) Representative NTA plots showing the size distribution and total number of EVs isolated from the same volume of conditioned medium of EGFR‐KO 5–8F and S18 cells. (c) Western blot analysis of phospho‐STAT3, STAT3, phospho‐AKT, AKT, phospho‐ERK1/2, and ERK1/2 in EVs‐treated, EGFR‐KO H‐EVs‐treated, EGFR overexpressed, and shEGFR transfected 6–10B and S26 cells. (d) Intracellular ROS levels in L‐EVs‐, H‐EVs‐ or EGFR‐KO H‐EVs‐treated, EGFR overexpressed, and shEGFR transfected 6–10B and S26 cells by fluorescence microplate reader. (e) Intracellular ROS levels in H‐EVs‐treated 6–10B and S26 cells pretreated with DMSO control, SH‐4‐54, SCH772984, or AZD5363. (f) Intracellular ROS levels in EGFR overexpressed 6–10B and S26 cells pretreated with DMSO control, SH‐4‐54, SCH772984, or AZD5363. (g) Western blot analysis of Ki‐67, PTEN, Bcl‐2, and Bax in L‐EVs‐, H‐EVs‐, or EGFR‐KO H‐EVs‐treated, EGFR overexpressed, and shEGFR 6–10B and S26 cells. (h) Clone formation capacity of H‐EVs‐treated, EGFR‐KO H‐EVs‐treated, EGFR overexpressed, ROS inhibited, shEGFR, AZD5363 treated, H‐EVs and AZD5363 co‐treated, and EGFR overexpression combined with AZD5363 treated 6–10B and S26 cells, respectively. (i) Wound healing assay showing cell migration of H‐EVs‐treated, EGFR‐KO H‐EVs‐treated, EGFR overexpressed, ROS inhibited, shEGFR,AZD5363 treated, H‐EVs and AZD5363 co‐treated, and EGFR overexpression combined with AZD5363 treated 6–10B and S26 cells. (j) Cell invasion of H‐EVs‐treated, EGFR‐KO H‐EVs‐treated, EGFR overexpressed, ROS inhibited, shEGFR, AZD5363 treated, H‐EVs and AZD5363 co‐treated, and EGFR overexpression combined with AZD5363 treated 6–10B and S26 cells using a Matrigel invasion assay. Experiments were performed in triplicate. Data are presented as mean ± SD ( : P
    Figure Legend Snippet: H‐EVs‐induced EGFR expression down‐regulates ROS via the PI3K/AKT pathway to promote metastasis of NPC cells. (a) Western blot analysis showing the presence of CD9, CD63, ALIX, and TSG101 and absence of GAPDH in EGFR‐KO EVs derived from the conditioned medium of EGFR‐KO 5–8F and S18 cells. (b) Representative NTA plots showing the size distribution and total number of EVs isolated from the same volume of conditioned medium of EGFR‐KO 5–8F and S18 cells. (c) Western blot analysis of phospho‐STAT3, STAT3, phospho‐AKT, AKT, phospho‐ERK1/2, and ERK1/2 in EVs‐treated, EGFR‐KO H‐EVs‐treated, EGFR overexpressed, and shEGFR transfected 6–10B and S26 cells. (d) Intracellular ROS levels in L‐EVs‐, H‐EVs‐ or EGFR‐KO H‐EVs‐treated, EGFR overexpressed, and shEGFR transfected 6–10B and S26 cells by fluorescence microplate reader. (e) Intracellular ROS levels in H‐EVs‐treated 6–10B and S26 cells pretreated with DMSO control, SH‐4‐54, SCH772984, or AZD5363. (f) Intracellular ROS levels in EGFR overexpressed 6–10B and S26 cells pretreated with DMSO control, SH‐4‐54, SCH772984, or AZD5363. (g) Western blot analysis of Ki‐67, PTEN, Bcl‐2, and Bax in L‐EVs‐, H‐EVs‐, or EGFR‐KO H‐EVs‐treated, EGFR overexpressed, and shEGFR 6–10B and S26 cells. (h) Clone formation capacity of H‐EVs‐treated, EGFR‐KO H‐EVs‐treated, EGFR overexpressed, ROS inhibited, shEGFR, AZD5363 treated, H‐EVs and AZD5363 co‐treated, and EGFR overexpression combined with AZD5363 treated 6–10B and S26 cells, respectively. (i) Wound healing assay showing cell migration of H‐EVs‐treated, EGFR‐KO H‐EVs‐treated, EGFR overexpressed, ROS inhibited, shEGFR,AZD5363 treated, H‐EVs and AZD5363 co‐treated, and EGFR overexpression combined with AZD5363 treated 6–10B and S26 cells. (j) Cell invasion of H‐EVs‐treated, EGFR‐KO H‐EVs‐treated, EGFR overexpressed, ROS inhibited, shEGFR, AZD5363 treated, H‐EVs and AZD5363 co‐treated, and EGFR overexpression combined with AZD5363 treated 6–10B and S26 cells using a Matrigel invasion assay. Experiments were performed in triplicate. Data are presented as mean ± SD ( : P

    Techniques Used: Expressing, Western Blot, Derivative Assay, Isolation, Transfection, Fluorescence, Over Expression, Wound Healing Assay, Migration, Invasion Assay

    19) Product Images from "Expression of LRIG1, a Negative Regulator of EGFR, Is Dynamically Altered during Different Stages of Gastric Carcinogenesis"

    Article Title: Expression of LRIG1, a Negative Regulator of EGFR, Is Dynamically Altered during Different Stages of Gastric Carcinogenesis

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2018.08.006

    Immunohistochemistry (IHC) analysis of leucine-rich repeats and immunoglobulin-like domains protein (LRIG)-1 and epidermal growth factor receptor (EGFR) expression in human gastric preneoplastic and cancer tissues. LRIG1 and EGFR were detected in human gastric preneoplastic and cancer tissues using IHC with indicated antibodies. A and B: In a spasmolytic polypeptide–expressing metaplastic (SPEM) region ( A ), LRIG1 was expressed strongly, whereas EGFR expression was low along SPEM glands ( arrowheads ). C–E: Decreased LRIG1 expression ( C and D ; yellow arrowheads ) and low EGFR gland expression ( C and D ; black arrowheads ) are observed in the intestinal metaplastic (IM) region, and LRIG1 is absent in cancer tissues ( E ). D and F: Strong EGFR expression in the IM region ( D , yellow arrowheads ) and in cancer lesions ( F ) is shown. Samples from five patients were used in these experiments G: LRIG1- or EGFR-positive cells were counted from 10 microscopic fields. Statistical analyses were performed using JMP software version 4. Data are expressed as means ± SEM. ∗ P
    Figure Legend Snippet: Immunohistochemistry (IHC) analysis of leucine-rich repeats and immunoglobulin-like domains protein (LRIG)-1 and epidermal growth factor receptor (EGFR) expression in human gastric preneoplastic and cancer tissues. LRIG1 and EGFR were detected in human gastric preneoplastic and cancer tissues using IHC with indicated antibodies. A and B: In a spasmolytic polypeptide–expressing metaplastic (SPEM) region ( A ), LRIG1 was expressed strongly, whereas EGFR expression was low along SPEM glands ( arrowheads ). C–E: Decreased LRIG1 expression ( C and D ; yellow arrowheads ) and low EGFR gland expression ( C and D ; black arrowheads ) are observed in the intestinal metaplastic (IM) region, and LRIG1 is absent in cancer tissues ( E ). D and F: Strong EGFR expression in the IM region ( D , yellow arrowheads ) and in cancer lesions ( F ) is shown. Samples from five patients were used in these experiments G: LRIG1- or EGFR-positive cells were counted from 10 microscopic fields. Statistical analyses were performed using JMP software version 4. Data are expressed as means ± SEM. ∗ P

    Techniques Used: Immunohistochemistry, Expressing, Software

    Effects of leucine-rich repeats and immunoglobulin-like domains protein (LRIG)-1 knockdown on tumorigenesis in the SNU638 gastric cancer cell line. A: LRIG1 was depleted using pGIPZ-based shRNA (sh46 and sh71). Levels of LRIG1, epidermal growth factor receptor (EGFR), phospho-EGFR, and epithelial to mesenchymal markers [E-cadherin, occludin, N-cadherin, vimentin, zinc finger E box–binding homeobox (ZEB)-1/T-cell factor (TCF)-3, and anti–zinc finger protein SNAI1 (SNAIL)] were detected by Western blot analysis using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a loading control. B: Cell proliferation was assessed with cell counting over 5 days. C: Cell cycle was examined with flow cytometry. D: Representative image of invasion and migration analyzed using Boyden chambers. E: Quantitative-analytic data from four independent experiments. Data are expressed as means ± SEM. n = 3 ( C ); n = 4 ( A , B , and D ). ∗ P
    Figure Legend Snippet: Effects of leucine-rich repeats and immunoglobulin-like domains protein (LRIG)-1 knockdown on tumorigenesis in the SNU638 gastric cancer cell line. A: LRIG1 was depleted using pGIPZ-based shRNA (sh46 and sh71). Levels of LRIG1, epidermal growth factor receptor (EGFR), phospho-EGFR, and epithelial to mesenchymal markers [E-cadherin, occludin, N-cadherin, vimentin, zinc finger E box–binding homeobox (ZEB)-1/T-cell factor (TCF)-3, and anti–zinc finger protein SNAI1 (SNAIL)] were detected by Western blot analysis using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a loading control. B: Cell proliferation was assessed with cell counting over 5 days. C: Cell cycle was examined with flow cytometry. D: Representative image of invasion and migration analyzed using Boyden chambers. E: Quantitative-analytic data from four independent experiments. Data are expressed as means ± SEM. n = 3 ( C ); n = 4 ( A , B , and D ). ∗ P

    Techniques Used: shRNA, Binding Assay, Western Blot, Cell Counting, Flow Cytometry, Cytometry, Migration

    Effects of leucine-rich repeats and immunoglobulin-like domains protein (LRIG)-1 knockdown on SNU638 xenografts and Kaplan-Meier survival curves of relapse-free survival (RFS) of gastric cancer patients with low or high LRIG1 mRNA expression. SNU638 cells, with or without vector transfection, were injected s.c. into the dorsal flank of 7-week-old male athymic nude mice to establish primary tumors. A: Tumor volumes were measured using calipers. B: Extent of tumor growth was evaluated in vivo . Dotted circles represent tumor mass. C: Histopathologic analysis of gastric tumor masses from mouse xenograft model. Hematoxylin and eosin (H E) staining was performed to investigate epidermal growth factor receptor (EGFR) and phospho (p)-EGFR (Tyr1068) expression in tumor tissues from xenograft mice. Boxed areas are shown at higher magnification to the right. EGFR- and pEGFR-positive cells were counted from five microscopic fields. Statistical analyses were performed using JMP software version 4. D: Kaplan-Meier curves show RFS in gastric cancer patients, monitored for 150 months, based on high or low tumor LRIG1 mRNA expression levels. Red, patients with expression levels above the median; black, patients with expression levels below the median. Patients with low expression levels had a lower probability of RFS over time. Data are expressed as means ± SEM. n = 4 mice per treatment. ∗ P
    Figure Legend Snippet: Effects of leucine-rich repeats and immunoglobulin-like domains protein (LRIG)-1 knockdown on SNU638 xenografts and Kaplan-Meier survival curves of relapse-free survival (RFS) of gastric cancer patients with low or high LRIG1 mRNA expression. SNU638 cells, with or without vector transfection, were injected s.c. into the dorsal flank of 7-week-old male athymic nude mice to establish primary tumors. A: Tumor volumes were measured using calipers. B: Extent of tumor growth was evaluated in vivo . Dotted circles represent tumor mass. C: Histopathologic analysis of gastric tumor masses from mouse xenograft model. Hematoxylin and eosin (H E) staining was performed to investigate epidermal growth factor receptor (EGFR) and phospho (p)-EGFR (Tyr1068) expression in tumor tissues from xenograft mice. Boxed areas are shown at higher magnification to the right. EGFR- and pEGFR-positive cells were counted from five microscopic fields. Statistical analyses were performed using JMP software version 4. D: Kaplan-Meier curves show RFS in gastric cancer patients, monitored for 150 months, based on high or low tumor LRIG1 mRNA expression levels. Red, patients with expression levels above the median; black, patients with expression levels below the median. Patients with low expression levels had a lower probability of RFS over time. Data are expressed as means ± SEM. n = 4 mice per treatment. ∗ P

    Techniques Used: Expressing, Plasmid Preparation, Transfection, Injection, Mouse Assay, In Vivo, Staining, Software

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

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

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

    Immunofluorescence:

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

    Staining:

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

    Electrophoresis:

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

    Real-time Polymerase Chain Reaction:

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

    Amplification:

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

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


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

    Journal: Experimental and Therapeutic Medicine

    Article Title: ABCG1 as a potential oncogene in lung cancer

    doi: 10.3892/etm.2017.4393

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

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

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

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

    Journal: Cancer prevention research (Philadelphia, Pa.)

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

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

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

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

    Techniques: Immunohistochemistry, Inhibition, Expressing, Staining

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

    Journal: Nature Communications

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

    doi: 10.1038/s41467-018-05368-1

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

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

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

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

    Journal: BMC Cancer

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

    doi: 10.1186/s12885-015-1401-z

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

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

    Techniques: Activation Assay, Western Blot, Expressing, Immunoprecipitation