Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: The c.1253 T > C (p.I418T) mutation has no effect on ERBB3 expression and interaction with ERBB2, whereas the c.3182dupA (p.N1061Kfs*16) mutation produces a novel truncated protein. a Western blotting results using anti-Myc antibody to detect ERBB3 in lysates of HEK293T cells transfected with 2 μg empty vector (EV) or WT, c.1253 T > C (M1), or c.3182dupA (M2) plasmids. b Results of co-immunoprecipitation to detect the interaction between ERBB2 and WT or I418T (M1) mutant ERBB3

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Mutagenesis, Expressing, Western Blot, Transfection, Plasmid Preparation, Immunoprecipitation

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: ERBB3 variants lack the capacity to activate PI3K/AKT and ERK signaling pathways. a Immunoblot analysis was performed using indicated antibodies to determine the effects of WT or mutant ERBB3 on PI3K/AKT and ERK pathway activation. To induce protein phosphorylation, HEK293T cells were treated with 10 ng/ml NRG-1β for 30 min after transfection with empty vector (EV), WT, M1, M2, or M3 (V104 L) plasmids. b–e Quantitative analysis of p-ERK, p-AKT, p-ERBB2, and p–ERBB3 expression. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. WT

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Protein-Protein interactions, Western Blot, Mutagenesis, Activation Assay, Phospho-proteomics, Transfection, Plasmid Preparation, Expressing

Journal: Molecular Medicine Reports

Article Title: EDIL3 knockdown inhibits retinal angiogenesis through the induction of cell cycle arrest in vitro

doi: 10.3892/mmr.2017.7122

Figure Lengend Snippet: Silencing EDIL3 expression interferes with the expression of cell cycle-regulatory proteins and EGFR signaling. HRECs were transfected with siEDIL3 or scramble siRNA and the expression of proteins of interest was detected using western blot analysis. (A) Silencing EDIL3 expression suppressed the protein expression of cyclin D1 and cyclin E1, whereas it had no effect on cyclin B1 expression. (B) Silencing EDIL3 expression appeared to inhibit the phosphorylation of EGFR, Src and ERK in HRECs in vitro . EDIL3, epidermal growth factor-like repeat and discoidin I-like domain-containing protein 3; EGFR, epidermal growth factor receptor; ERK, extracellular signal-regulated kinase; HRECs, human retinal endothelial cells; p-, phosphorylated; si, small interfering.

Article Snippet: Antibodies against ERK (Catalog: 4695), phosphorylated (p)-ERK (Catalog: 4370), EGFR (Catalog: 2085), p-EGFR (Catalog: 2244), Src (Catalog: 2109) and p-Src (Catalog: 12432) were purchased from CST Biological Reagents Company Limited (Shanghai, China).

Techniques: Expressing, Transfection, Western Blot, Phospho-proteomics, In Vitro

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: The c.1253 T > C (p.I418T) mutation has no effect on ERBB3 expression and interaction with ERBB2, whereas the c.3182dupA (p.N1061Kfs*16) mutation produces a novel truncated protein. a Western blotting results using anti-Myc antibody to detect ERBB3 in lysates of HEK293T cells transfected with 2 μg empty vector (EV) or WT, c.1253 T > C (M1), or c.3182dupA (M2) plasmids. b Results of co-immunoprecipitation to detect the interaction between ERBB2 and WT or I418T (M1) mutant ERBB3

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Mutagenesis, Expressing, Western Blot, Transfection, Plasmid Preparation, Immunoprecipitation

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: ERBB3 variants lack the capacity to activate PI3K/AKT and ERK signaling pathways. a Immunoblot analysis was performed using indicated antibodies to determine the effects of WT or mutant ERBB3 on PI3K/AKT and ERK pathway activation. To induce protein phosphorylation, HEK293T cells were treated with 10 ng/ml NRG-1β for 30 min after transfection with empty vector (EV), WT, M1, M2, or M3 (V104 L) plasmids. b–e Quantitative analysis of p-ERK, p-AKT, p-ERBB2, and p–ERBB3 expression. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. WT

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Protein-Protein interactions, Western Blot, Mutagenesis, Activation Assay, Phospho-proteomics, Transfection, Plasmid Preparation, Expressing

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: ERBB3 variants lack the capacity to activate PI3K/AKT and ERK signaling pathways. a Immunoblot analysis was performed using indicated antibodies to determine the effects of WT or mutant ERBB3 on PI3K/AKT and ERK pathway activation. To induce protein phosphorylation, HEK293T cells were treated with 10 ng/ml NRG-1β for 30 min after transfection with empty vector (EV), WT, M1, M2, or M3 (V104 L) plasmids. b–e Quantitative analysis of p-ERK, p-AKT, p-ERBB2, and p–ERBB3 expression. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. WT

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Protein-Protein interactions, Western Blot, Mutagenesis, Activation Assay, Phospho-proteomics, Transfection, Plasmid Preparation, Expressing

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: Clinical features of patients with biallelic ERBB3 mutations

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Variant Assay, Infection

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: Genomic DNA sequencing of the pedigree. a Pedigree of the patient. b The data analysis algorithm used for filtering all single nucleotide variants identified using trio-based, whole exome sequencing, with the number of remaining variants after each filtering step. On filtering and prioritization, compound heterozygous variants of the ERBB3 gene were identified as the top candidate. MAF, minor allele frequency. c Sanger sequencing confirmed the compound heterozygous variants, c.1253 T > C;p.I418T and c.3182dupA;p.N1061Kfs*16, in the patient. Red arrows indicate variant bases. d Distribution of loss-of-function germline mutations in the ERBB3 protein

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: DNA Sequencing, Sequencing, Variant Assay

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: In silico analysis of the p.Ile418Thr variant. a Alignment of amino acid sequences among various species; the position of the mutant residue within the highly conserved region is indicated in red. b , c Homology models of the WT ERBB3 ( b ) and p.Ile418Thr mutant ERBB3 ( c ) N-terminal tails of the extracellular domain. Residue at position 418 is shown in yellow

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: In Silico, Variant Assay, Mutagenesis, Residue

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: The c.1253 T > C (p.I418T) mutation has no effect on ERBB3 expression and interaction with ERBB2, whereas the c.3182dupA (p.N1061Kfs*16) mutation produces a novel truncated protein. a Western blotting results using anti-Myc antibody to detect ERBB3 in lysates of HEK293T cells transfected with 2 μg empty vector (EV) or WT, c.1253 T > C (M1), or c.3182dupA (M2) plasmids. b Results of co-immunoprecipitation to detect the interaction between ERBB2 and WT or I418T (M1) mutant ERBB3

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Mutagenesis, Expressing, Western Blot, Transfection, Plasmid Preparation, Immunoprecipitation

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: ERBB3 variants lack the capacity to activate PI3K/AKT and ERK signaling pathways. a Immunoblot analysis was performed using indicated antibodies to determine the effects of WT or mutant ERBB3 on PI3K/AKT and ERK pathway activation. To induce protein phosphorylation, HEK293T cells were treated with 10 ng/ml NRG-1β for 30 min after transfection with empty vector (EV), WT, M1, M2, or M3 (V104 L) plasmids. b–e Quantitative analysis of p-ERK, p-AKT, p-ERBB2, and p–ERBB3 expression. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. WT

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Protein-Protein interactions, Western Blot, Mutagenesis, Activation Assay, Phospho-proteomics, Transfection, Plasmid Preparation, Expressing

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: ERBB3 variants lack the capacity to activate PI3K/AKT and ERK signaling pathways. a Immunoblot analysis was performed using indicated antibodies to determine the effects of WT or mutant ERBB3 on PI3K/AKT and ERK pathway activation. To induce protein phosphorylation, HEK293T cells were treated with 10 ng/ml NRG-1β for 30 min after transfection with empty vector (EV), WT, M1, M2, or M3 (V104 L) plasmids. b–e Quantitative analysis of p-ERK, p-AKT, p-ERBB2, and p–ERBB3 expression. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. WT

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Protein-Protein interactions, Western Blot, Mutagenesis, Activation Assay, Phospho-proteomics, Transfection, Plasmid Preparation, Expressing

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: Clinical features of patients with biallelic ERBB3 mutations

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Variant Assay, Infection

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: Genomic DNA sequencing of the pedigree. a Pedigree of the patient. b The data analysis algorithm used for filtering all single nucleotide variants identified using trio-based, whole exome sequencing, with the number of remaining variants after each filtering step. On filtering and prioritization, compound heterozygous variants of the ERBB3 gene were identified as the top candidate. MAF, minor allele frequency. c Sanger sequencing confirmed the compound heterozygous variants, c.1253 T > C;p.I418T and c.3182dupA;p.N1061Kfs*16, in the patient. Red arrows indicate variant bases. d Distribution of loss-of-function germline mutations in the ERBB3 protein

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: DNA Sequencing, Sequencing, Variant Assay

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: In silico analysis of the p.Ile418Thr variant. a Alignment of amino acid sequences among various species; the position of the mutant residue within the highly conserved region is indicated in red. b , c Homology models of the WT ERBB3 ( b ) and p.Ile418Thr mutant ERBB3 ( c ) N-terminal tails of the extracellular domain. Residue at position 418 is shown in yellow

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: In Silico, Variant Assay, Mutagenesis, Residue

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: The c.1253 T > C (p.I418T) mutation has no effect on ERBB3 expression and interaction with ERBB2, whereas the c.3182dupA (p.N1061Kfs*16) mutation produces a novel truncated protein. a Western blotting results using anti-Myc antibody to detect ERBB3 in lysates of HEK293T cells transfected with 2 μg empty vector (EV) or WT, c.1253 T > C (M1), or c.3182dupA (M2) plasmids. b Results of co-immunoprecipitation to detect the interaction between ERBB2 and WT or I418T (M1) mutant ERBB3

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Mutagenesis, Expressing, Western Blot, Transfection, Plasmid Preparation, Immunoprecipitation

Journal: Orphanet Journal of Rare Diseases

Article Title: Biallelic ERBB3 loss-of-function variants are associated with a novel multisystem syndrome without congenital contracture

doi: 10.1186/s13023-019-1241-z

Figure Lengend Snippet: ERBB3 variants lack the capacity to activate PI3K/AKT and ERK signaling pathways. a Immunoblot analysis was performed using indicated antibodies to determine the effects of WT or mutant ERBB3 on PI3K/AKT and ERK pathway activation. To induce protein phosphorylation, HEK293T cells were treated with 10 ng/ml NRG-1β for 30 min after transfection with empty vector (EV), WT, M1, M2, or M3 (V104 L) plasmids. b–e Quantitative analysis of p-ERK, p-AKT, p-ERBB2, and p–ERBB3 expression. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. WT

Article Snippet: Antibodies against ERBB2 (#2242), phosphorylated (p-)ERBB2 (Tyr1248) (#2247), ERBB3 (C-terminus) (#12708), p-ERBB3 (Tyr1289) (#4791), ERK (#4695), p-ERK (#4370), AKT (#4685), p-AKT (#4060), GFP (#2555), Myc (#2272), and GAPDH (#5174) were purchased from Cell Signaling Technology.

Techniques: Protein-Protein interactions, Western Blot, Mutagenesis, Activation Assay, Phospho-proteomics, Transfection, Plasmid Preparation, Expressing