Journal: International Journal of Clinical and Experimental Pathology

Article Title: FOXO3a expression and its diagnostic value in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: The expression of FOXO3a in normal tissues and tumor tissues

Article Snippet: Subsequently, we used the avidin-biotin-peroxidase method to stain the tissues, and we also used the anti-FOXO3a rabbit monoclonal antibody (CST D19A7; dilution 1:200) as primary antibody.

Techniques: Expressing, Staining

Journal: International Journal of Clinical and Experimental Pathology

Article Title: FOXO3a expression and its diagnostic value in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Normal pancreatic tissue and PDAC were stained with antibodies (Ab) specific to FOXO3a. (A, B) FOXO3a over-expression in PDAC. (C, D) Tumor tissue stained with PBS. (E, F) FOXO3a negative staining in normal pancreatic tissue (A, C, E: ×200, B, D, F: ×400).

Article Snippet: Subsequently, we used the avidin-biotin-peroxidase method to stain the tissues, and we also used the anti-FOXO3a rabbit monoclonal antibody (CST D19A7; dilution 1:200) as primary antibody.

Techniques: Staining, Over Expression, Negative Staining

Journal: International Journal of Clinical and Experimental Pathology

Article Title: FOXO3a expression and its diagnostic value in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: The relationship between FOXO3a expression in tumor tissues and clinicopathologic features

Article Snippet: Subsequently, we used the avidin-biotin-peroxidase method to stain the tissues, and we also used the anti-FOXO3a rabbit monoclonal antibody (CST D19A7; dilution 1:200) as primary antibody.

Techniques: Expressing

Journal: International Journal of Clinical and Experimental Pathology

Article Title: FOXO3a expression and its diagnostic value in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Univariate analysis of the overall survival of patients with PDAC

Article Snippet: Subsequently, we used the avidin-biotin-peroxidase method to stain the tissues, and we also used the anti-FOXO3a rabbit monoclonal antibody (CST D19A7; dilution 1:200) as primary antibody.

Techniques: Expressing, Adjuvant

Journal: International Journal of Clinical and Experimental Pathology

Article Title: FOXO3a expression and its diagnostic value in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Kaplan-Meier survival curves of FOXO3a expression in PDAC (log-rank test). A. All patients (P=0.013). B. Patients without postoperative adjuvant therapy (P<0.001).

Article Snippet: Subsequently, we used the avidin-biotin-peroxidase method to stain the tissues, and we also used the anti-FOXO3a rabbit monoclonal antibody (CST D19A7; dilution 1:200) as primary antibody.

Techniques: Expressing, Adjuvant

Journal: International Journal of Clinical and Experimental Pathology

Article Title: FOXO3a expression and its diagnostic value in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Multivariate analysis of overall survival

Article Snippet: Subsequently, we used the avidin-biotin-peroxidase method to stain the tissues, and we also used the anti-FOXO3a rabbit monoclonal antibody (CST D19A7; dilution 1:200) as primary antibody.

Techniques: Expressing, Adjuvant

Journal: International Journal of Clinical and Experimental Pathology

Article Title: FOXO3a expression and its diagnostic value in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Multivariate Analysis of Overall Survival of patients without postoperative adjuvant therapy

Article Snippet: Subsequently, we used the avidin-biotin-peroxidase method to stain the tissues, and we also used the anti-FOXO3a rabbit monoclonal antibody (CST D19A7; dilution 1:200) as primary antibody.

Techniques: Adjuvant, Expressing

Journal: Human Mutation

Article Title: A Comprehensive Functional Analysis of NTRK1 Missense Mutations Causing Hereditary Sensory and Autonomic Neuropathy Type IV (HSAN IV)

doi: 10.1002/humu.23123

Figure Lengend Snippet: TRKA structure, signaling, and identification of novel NTRK1 mutations in patients with HSAN IV. A : NTRK1 located on chromosome 1q21‐q22, shown by the gray bar on the ideogram, consists of 17 exons and spans region of 25 kb. Inclusion of all 17 exons encodes for the neuronal specific isoform of TRKA. B : The extracellular domain of TRKA consists of a number of structural domains: a leucine rich repeat, flanked on either side by a cysteine rich domain, and two immunoglobulin‐like domains. The majority of the intracellular domain comprises the kinase domain. C : Signaling through TRKA is initiated through NGF binding, TRKA dimerization, and autophosphorylation of Y676, Y680, and Y681. Subsequent phosphorylation of Y496 and Y791 leads to activation of downstream signaling cascades which drive neuronal differentiation and survival. D : Seven individuals were ascertained with a standard HSAN IV phenotype. The identified mutations were p.G517E, p.G522E, p.L657P, p.I699T, p.C752S, p.C763S, and p.R771C. Only the exons that code for the kinase domain are shown. TKD, tyrosine kinase domain.

Article Snippet: Total TRKA levels in the lysates were also measured using PathScan® Total TRKA Sandwich ELISA kit (CST).

Techniques: Binding Assay, Phospho-proteomics, Activation Assay

Journal: Human Mutation

Article Title: A Comprehensive Functional Analysis of NTRK1 Missense Mutations Causing Hereditary Sensory and Autonomic Neuropathy Type IV (HSAN IV)

doi: 10.1002/humu.23123

Figure Lengend Snippet: Expression, processing, and localization of wild‐type and mutant TRKA. A : Expression of C‐terminal GFP tagged wild‐type and mutant TRKA constructs in transfected HEK‐293 cells. The lanes “+” and “−” represent lysates treated with/without PNGase respectively. Lamin B1 was included as a loading control. Representative image of n = 3 is shown. WT, wild‐type; EV, empty vector. B : Quantification of relative levels of fully glycosylated 170 kDa TRKA‐GFP protein to the levels of partially glycosylated 130 kDa TRKA‐GFP protein. C : Localization of TRKA‐GFP wild‐type and mutant proteins in transfected HEK‐293 cells. The cells were stained against GFP (green) and the plasma membrane marker Na + /K + ATPase (red). Wild‐type TRKA and all the mutants show membrane expression. D : Relative levels of GFP at the membrane compared with the levels of GFP in the cytoplasm. Graph represents mean values of n = 3, with at least 15 cells in each repeat, and error bars represent standard error of the mean. Statistical differences are indicated as * P < 0.05, ** P < 0.01 (one‐way ANOVA, followed by Student's t ‐test using a Bonferroni adjusted P value).

Article Snippet: Total TRKA levels in the lysates were also measured using PathScan® Total TRKA Sandwich ELISA kit (CST).

Techniques: Expressing, Mutagenesis, Construct, Transfection, Control, Plasmid Preparation, Staining, Clinical Proteomics, Membrane, Marker

Journal: Human Mutation

Article Title: A Comprehensive Functional Analysis of NTRK1 Missense Mutations Causing Hereditary Sensory and Autonomic Neuropathy Type IV (HSAN IV)

doi: 10.1002/humu.23123

Figure Lengend Snippet: Relative Y680/681 and Y496 phosphorylation of mutant TRKA. A : Basal level of wild‐type and mutant TRKA‐GFP autophosphorylation in transfected untreated HeLa cells. Response to wild‐type and mutant TRKA‐GFP protein to 15 min stimulation with 100 ng/ml NGF. B : Basal level of Y496 phosphorylation in wild‐type and mutant TRKA proteins in untreated cells and after NGF stimulation. The bar graphs in A and B represents the mean values of n = 3 and error bars represents standard error of the mean. Statistical differences between induced phosphorylation of wild‐type and mutant TRKA are indicated as ** P < 0.01 or *** P < 0.001 (one‐way ANOVA, followed by Student's t ‐test using a Bonferroni adjusted P value). WT, wild‐type; EV, empty vector.

Article Snippet: Total TRKA levels in the lysates were also measured using PathScan® Total TRKA Sandwich ELISA kit (CST).

Techniques: Phospho-proteomics, Mutagenesis, Transfection, Plasmid Preparation

Journal: Human Mutation

Article Title: A Comprehensive Functional Analysis of NTRK1 Missense Mutations Causing Hereditary Sensory and Autonomic Neuropathy Type IV (HSAN IV)

doi: 10.1002/humu.23123

Figure Lengend Snippet: Effect of mutations on induction of PLCγ pathway. A : HeLa cells were transfected with wild‐type and mutant TRKA constructs and loaded with rhodamine‐3 and response to NGF was monitored. Ionomycin was used as the positive control. Only cells that did not spontaneously fluoresce and had an ionomycin response were used for analysis. WT, wild‐type. B : No significant increase in calcium levels was observed after NGF stimulation in p.G517E transfected cells. C : There was an increase in fluorescence in p.C752S transfected cells which peaked at t = 208. D : NGF stimulation on cells transfected with p.C763S caused an increase in fluorescence but the maximum value was reduced.

Article Snippet: Total TRKA levels in the lysates were also measured using PathScan® Total TRKA Sandwich ELISA kit (CST).

Techniques: Transfection, Mutagenesis, Construct, Positive Control, Fluorescence

Journal: Human Mutation

Article Title: A Comprehensive Functional Analysis of NTRK1 Missense Mutations Causing Hereditary Sensory and Autonomic Neuropathy Type IV (HSAN IV)

doi: 10.1002/humu.23123

Figure Lengend Snippet: Summary of Functional Characterization of New TRKA Mutations

Article Snippet: Total TRKA levels in the lysates were also measured using PathScan® Total TRKA Sandwich ELISA kit (CST).

Techniques: Functional Assay, Mutagenesis, Glycoproteomics, Membrane, Expressing, Activation Assay

Journal: Journal of neurochemistry

Article Title: Molecular Mechanisms Underlying Protective Effects of Quercetin Against Mitochondrial Dysfunction and Progressive Dopaminergic Neurodegeneration in Cell Culture and MitoPark Transgenic Mouse Models of Parkinson’s Disease

doi: 10.1111/jnc.14033

Figure Lengend Snippet: A, MN9D cells were treated with varying concentrations of quercetin (0.3–30 µM) for 24 h and cell lysates were prepared and subjected to Western blot analysis. Representative immunoblots of phospho-CREB (S133), total CREB, and BDNF (n=2–3). B, MN9D cells were treated with 10 and 30 µM quercetin for 24 h and real-time RT-PCR analysis of BDNF mRNA level was performed (n=9). 18S rRNA served as an internal control. Values represented as a percentage of the activity of control and are expressed as means ± SEM of three independent experiments performed in triplicate (***p<0.001 comparing control and quercetin-treated samples).

Article Snippet: Antibodies to phospho PKD1 (p-S744/748 (Cell Signaling Technology (CST) Cat# 2054S RRID:AB_2172539) and p-S916 (CST Cat# 2051S RRID:AB_330841)), phospho-Akt (p-S473) (CST Cat# 4060 also 4060L, 4060S, 4060P RRID:AB_2315049), total Akt (CST Cat# 4691 also 4691L, 4691P, 4691S RRID:AB_915783), total CREB (CST Cat# 9197 also 9197S, 9197L RRID:AB_331277), and phospho-CREB (p-S133) (CST Cat# 9198 also 9198S, 9198L RRID:AB_2561044) were obtained from CST (Danvers, MA).

Techniques: Western Blot, Quantitative RT-PCR, Control, Activity Assay