Journal: PLoS ONE

Article Title: An Integrative Analysis to Identify Driver Genes in Esophageal Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0139808

Figure Lengend Snippet: GISTIC-defined peaks associated with altered expression in ESCC.

Article Snippet: ERBB2 and GRB7 proteins were detected using an anti-ERBB2 rabbit monoclonal antibody (Epitomics, Inc) at a 1:5000 dilution and an anti-GRB7 rabbit monoclonal antibody (Gene Tex International Corporation) at a 1:1000 dilution.

Techniques: Expressing, Amplification

Journal: PLoS ONE

Article Title: An Integrative Analysis to Identify Driver Genes in Esophageal Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0139808

Figure Lengend Snippet: Results of analysis on RNA interference screening data of Project Achilles.

Article Snippet: ERBB2 and GRB7 proteins were detected using an anti-ERBB2 rabbit monoclonal antibody (Epitomics, Inc) at a 1:5000 dilution and an anti-GRB7 rabbit monoclonal antibody (Gene Tex International Corporation) at a 1:1000 dilution.

Techniques: Virus

Journal: PLoS ONE

Article Title: An Integrative Analysis to Identify Driver Genes in Esophageal Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0139808

Figure Lengend Snippet: (a) Reductions in mRNA and protein levels of GRB7 at 48 hours after siRNA transfection in KYSE410 and TE4 cells. The results are the mean ± SD from 3 replicates of a single experiment. (b) GRB7 inactivation reduced proliferation of KYSE410 and TE4 cells. Cell growth was measured on days 2, 3, and 4 by MTT assay. Absorbance at day 0 was assigned a value of 1. The results are the mean ± SD from 6 replicates of a single experiment. (c) Migration and invasion assays using GRB7 -knockdown cells. Each bar represents the average of 3 measurements. (d) Inhibitory effects of siRNA targeting GRB7 in combination with trastuzumab. Cells were transfected with siRNA targeting GRB7 or negative control siRNA and treated with or without trastuzumab (0.1 and 1.0 μg/mL). Cells were then seeded in 96-well plates, and cell growth was monitored every 24 hours using MTT assays. Absorbance at day 0 was assigned a value of 1. The results are the mean ± SD from 6 replicates of a single experiment.

Article Snippet: ERBB2 and GRB7 proteins were detected using an anti-ERBB2 rabbit monoclonal antibody (Epitomics, Inc) at a 1:5000 dilution and an anti-GRB7 rabbit monoclonal antibody (Gene Tex International Corporation) at a 1:1000 dilution.

Techniques: Transfection, MTT Assay, Migration, Knockdown, Negative Control

Journal: PLoS ONE

Article Title: An Integrative Analysis to Identify Driver Genes in Esophageal Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0139808

Figure Lengend Snippet: (a) Analysis of GRB7 mRNA expression in tumor tissues and the corresponding normal mucosa by real-time RT-PCR. (b) Kaplan-Meier survival curves for ESCC patients according to GRB7 mRNA expression.

Article Snippet: ERBB2 and GRB7 proteins were detected using an anti-ERBB2 rabbit monoclonal antibody (Epitomics, Inc) at a 1:5000 dilution and an anti-GRB7 rabbit monoclonal antibody (Gene Tex International Corporation) at a 1:1000 dilution.

Techniques: Expressing, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: An Integrative Analysis to Identify Driver Genes in Esophageal Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0139808

Figure Lengend Snippet: Results of univariate and multivariate analysis of clinicopathlogical factors for 5-year overall survival in the validation set.

Article Snippet: ERBB2 and GRB7 proteins were detected using an anti-ERBB2 rabbit monoclonal antibody (Epitomics, Inc) at a 1:5000 dilution and an anti-GRB7 rabbit monoclonal antibody (Gene Tex International Corporation) at a 1:1000 dilution.

Techniques: Biomarker Discovery, Expressing

Journal: PloS one

Article Title: Novel nonphosphorylated peptides with conserved sequences selectively bind to Grb7 SH2 domain with affinity comparable to its phosphorylated ligand.

doi: 10.1371/journal.pone.0029902

Figure Lengend Snippet: Figure 1. The nonphosphorylated peptide 41 and the Grb7 SH2 domain are co-immunoprecipitated from transfected HEK293T cells. A) Expression of nonphosphorylated peptides. B) Interactions between nonphosphorylated peptides and the Grb7 SH2 domain. doi:10.1371/journal.pone.0029902.g001

Article Snippet: Materials The ABL_1 and Grb2 plasmids were gifts from Professor Ruibao Ren (Brandeis University, USA), and the Grb7 (BC006535) and Grb14 cDNA (BC053559) plasmids were from the Proteintech Group.

Techniques: Immunoprecipitation, Transfection, Expressing

Journal: PloS one

Article Title: Novel nonphosphorylated peptides with conserved sequences selectively bind to Grb7 SH2 domain with affinity comparable to its phosphorylated ligand.

doi: 10.1371/journal.pone.0029902

Figure Lengend Snippet: Figure 2. BRET assay of interactions between nonphosphorylated peptides and the Grb7 SH2 domain. A) Overlaid images of transfected EYFP-fused proteins in living HEK293T cells at 480 nm. B) Quantification of the BRET luminescence ratios. The direct luminescence ratios of 530/480 nm were on the top, and the relative luminescence ratios of 530/480 nm were on the bottom. The BRET reaction was initiated by the coelenterazine (5.9 mM). The statistical analysis was based on eighteen group ratios, * represented statistical significance with p,0.05 and ** represented statistical significance with p,0.01. doi:10.1371/journal.pone.0029902.g002

Article Snippet: Materials The ABL_1 and Grb2 plasmids were gifts from Professor Ruibao Ren (Brandeis University, USA), and the Grb7 (BC006535) and Grb14 cDNA (BC053559) plasmids were from the Proteintech Group.

Techniques: Bioluminescence Resonance Energy Transfer, Transfection

Journal: PloS one

Article Title: Novel nonphosphorylated peptides with conserved sequences selectively bind to Grb7 SH2 domain with affinity comparable to its phosphorylated ligand.

doi: 10.1371/journal.pone.0029902

Figure Lengend Snippet: Figure 3. The nonphosphorylated peptide has at least comparable binding affinity to the phosphorylated ligand pY (1180) for the Grb7 SH2 domain. A) Different concentrations of synthesized phosphorylated ligands pY (1180) (DEEYEpY(1180)MNRRR) from ErbB3 and the nonphosphorylated ligands 41-A (VAVGIPTQPTTSSEPSPPSNPPWDPGRV) from the nonphosphorylated ligand 41 were added respectively into the interacting complexes of Grb7 SH2 domain and peptide 41 (on the top). The bands were scanned and IC50 values were calculated (on the bottom). B)

Article Snippet: Materials The ABL_1 and Grb2 plasmids were gifts from Professor Ruibao Ren (Brandeis University, USA), and the Grb7 (BC006535) and Grb14 cDNA (BC053559) plasmids were from the Proteintech Group.

Techniques: Binding Assay, Synthesized

Journal: PloS one

Article Title: Novel nonphosphorylated peptides with conserved sequences selectively bind to Grb7 SH2 domain with affinity comparable to its phosphorylated ligand.

doi: 10.1371/journal.pone.0029902

Figure Lengend Snippet: Figure 4. Effect of the nonphosphorylated peptide 41 on the proliferation of the SK-BR-3 breast cancer cells. A) Protein expression. The endogenous Grb7 protein expression (left) and expression of the fusion protein of 41-mRLUC-36Flag (right) in the SK-BR-3 breast cancer cells were identified by western blot. B) SK-BR-3 breast cancer cell proliferation analysis. Double-factor variance analysis was performed to analyze the differences between the adjusted absorbance and different transfected cell groups, and between the adjusted absorbance and different time points. Statistically significant differences existed in both of groups (n = 10, p,0.001). doi:10.1371/journal.pone.0029902.g004

Article Snippet: Materials The ABL_1 and Grb2 plasmids were gifts from Professor Ruibao Ren (Brandeis University, USA), and the Grb7 (BC006535) and Grb14 cDNA (BC053559) plasmids were from the Proteintech Group.

Techniques: Expressing, Western Blot, Transfection