Journal: Cancer Management and Research

Article Title: DSG2 and c-MYC Interact to Regulate the Expression of ADAM17 and Promote the Development of Cervical Cancer

doi: 10.2147/CMAR.S456548

Figure Lengend Snippet: DSG2 is up-regulated in human cervical cancer and is associated with a poor prognosis. ( A ) Higher expression of DSG2 was found in cervical cancer samples than the normal tissues (based on TCGA database). ( B ) Kaplan–Meier plots of overall survival for cervical cancer samples with high/low DSG2 expression from the TCGA database.

Article Snippet: Primary antibodies against DSG2 (21,880-1-AP, 1:1000, Proteintech), ADAM17 (29,948-1-AP, 1:2000, Proteintech) and c-MYC (67,447-1-Ig, 1:5000, Proteintech) were used.

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: DSG2 and c-MYC Interact to Regulate the Expression of ADAM17 and Promote the Development of Cervical Cancer

doi: 10.2147/CMAR.S456548

Figure Lengend Snippet: Knockdown of DSG2 in cervical cancer cells. ( A ) The knockdown efficiency of the three groups of siRNA in HeLa cells was 84%, 86% and 28%, respectively. ( B and C ) The specificity and validity of the siRNA knockdown of DSG2 expression in HeLa and SiHa cells was verified by qPCR ( B ) and WB ( C ). *P < 0.05, **P < 0.01.

Article Snippet: Primary antibodies against DSG2 (21,880-1-AP, 1:1000, Proteintech), ADAM17 (29,948-1-AP, 1:2000, Proteintech) and c-MYC (67,447-1-Ig, 1:5000, Proteintech) were used.

Techniques: Knockdown, Expressing

Journal: Cancer Management and Research

Article Title: DSG2 and c-MYC Interact to Regulate the Expression of ADAM17 and Promote the Development of Cervical Cancer

doi: 10.2147/CMAR.S456548

Figure Lengend Snippet: Knockdown of DSG2 inhibited proliferation and migration of cervical cancer cells. ( A ) The proliferation of HeLa and SiHa cells after knockdown of DSG2 was measured using CCK-8 assay. ( B ) The effect of DSG2 knockdown on cervical cancer cell clone formation. ( C ) The migration ability of HeLa and SiHa cells after knockdown of DSG2 was measured using Transwell assay. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Primary antibodies against DSG2 (21,880-1-AP, 1:1000, Proteintech), ADAM17 (29,948-1-AP, 1:2000, Proteintech) and c-MYC (67,447-1-Ig, 1:5000, Proteintech) were used.

Techniques: Knockdown, Migration, CCK-8 Assay, Transwell Assay

Journal: Cancer Management and Research

Article Title: DSG2 and c-MYC Interact to Regulate the Expression of ADAM17 and Promote the Development of Cervical Cancer

doi: 10.2147/CMAR.S456548

Figure Lengend Snippet: ADAM17 is up-regulated in human cervical cancer and is associated with a poor prognosis. ( A ) The bioinformatics analysis of TCGA-CHOL dataset showed the positive expression correlation between DSG2 and ADAM17. ( B ) Higher expression of ADAM17 was found in cervical cancer samples than the normal tissues (based on TCGA database). ( C ) Kaplan–Meier plots of overall survival for cervical cancer samples with high/low ADAM17 expression from the TCGA database.

Article Snippet: Primary antibodies against DSG2 (21,880-1-AP, 1:1000, Proteintech), ADAM17 (29,948-1-AP, 1:2000, Proteintech) and c-MYC (67,447-1-Ig, 1:5000, Proteintech) were used.

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: DSG2 and c-MYC Interact to Regulate the Expression of ADAM17 and Promote the Development of Cervical Cancer

doi: 10.2147/CMAR.S456548

Figure Lengend Snippet: DSG2 regulates the expression of ADAM17 in cervical cancer. ( A ) The effect of DSG2 knockdown on ADAM17mRNA expression was detected by qPCR. ( B ) The effect of DSG2 knockdown on ADAM17 protein expression was detected by WB. ( C ) The interaction between DSG2 and c-MYC was detected by Co-IP assay. ***P < 0.001.

Article Snippet: Primary antibodies against DSG2 (21,880-1-AP, 1:1000, Proteintech), ADAM17 (29,948-1-AP, 1:2000, Proteintech) and c-MYC (67,447-1-Ig, 1:5000, Proteintech) were used.

Techniques: Expressing, Knockdown, Co-Immunoprecipitation Assay

Journal: Cancer Management and Research

Article Title: DSG2 and c-MYC Interact to Regulate the Expression of ADAM17 and Promote the Development of Cervical Cancer

doi: 10.2147/CMAR.S456548

Figure Lengend Snippet: DSG2 regulates cervical cancer development by interacting with c-MYC. ( A ) HeLa cells were transfected with pcDNA(3.1)-DSG2 overexpression plasmid, and the mRNA level of DSG2 was detected by qPCR. ( B ) HeLa cells were transfected with pcDNA(3.1) overexpression plasmid, and the protein level of DSG2 was detected by WB. ( C and D ) DSG2 overexpressed HeLa cells were treated with a C-MYC inhibitor (10,058-F4, 50 μM), and the proliferative activity and migration ability were detected by CCK-8 ( C ) and clonal formation assay ( D ). ( E ) DSG2 overexpressed HeLa cells were treated with a C-MYC inhibitor, and ADAM17 protein expression was detected by WB.*P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Primary antibodies against DSG2 (21,880-1-AP, 1:1000, Proteintech), ADAM17 (29,948-1-AP, 1:2000, Proteintech) and c-MYC (67,447-1-Ig, 1:5000, Proteintech) were used.

Techniques: Transfection, Over Expression, Plasmid Preparation, Activity Assay, Migration, CCK-8 Assay, Tube Formation Assay, Expressing

Journal: British journal of cancer

Article Title: Desmogleins as prognostic biomarkers in resected pancreatic ductal adenocarcinoma.

doi: 10.1038/bjc.2015.362

Figure Lengend Snippet: Figure 2. Variable expression of DSG1, DSG2 and DSG3 in normal pancreatic and PDAC tissue. Immunohistochemical staining of DSG1, DSG2 and DSG3 in exemplary normal pancreatic tissue as well as in exemplary PDAC cases classified as high or low expression, according to the staining intensity. It shows a variable expression pattern of DSG1 and DSG2 in normal pancreatic and PDAC tissue, whereas strong DSG3 expression is confined to PDAC tissue. 200-fold magnification. Scale bars, 50 mm.

Article Snippet: Immunohistochemical staining of 4 mm sections was done strictly following the manufacturer’s instructions on a Ventana Benchmark XT autostainer (Ventana Medical Systems, Oro Valley, AZ, USA) using an anti-DSG1 rabbit monoclonal primary antibody (clone EPR6766(B), Abcam, Cambridge, UK; dilution 1 : 400), an anti-DSG2 rabbit polyclonal primary antibody (Atlas Antibodies, Stockholm, Sweden; dilution 1 : 350) and an antiDSG3 mouse monoclonal primary antibody (clone 7B9, Abcam; dilution 1 : 150).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: British journal of cancer

Article Title: Desmogleins as prognostic biomarkers in resected pancreatic ductal adenocarcinoma.

doi: 10.1038/bjc.2015.362

Figure Lengend Snippet: Figure 3. The DSG3 expression correlates with inferior survival of PDAC patients. Univariate correlation analysis (Kaplan–Meier curves and log-rank tests) of (A) DSG1, (B) DSG2 and (C) DSG3 expression and postoperative survival in resected PDAC patients. Crossed lines indicate censored cases.

Article Snippet: Immunohistochemical staining of 4 mm sections was done strictly following the manufacturer’s instructions on a Ventana Benchmark XT autostainer (Ventana Medical Systems, Oro Valley, AZ, USA) using an anti-DSG1 rabbit monoclonal primary antibody (clone EPR6766(B), Abcam, Cambridge, UK; dilution 1 : 400), an anti-DSG2 rabbit polyclonal primary antibody (Atlas Antibodies, Stockholm, Sweden; dilution 1 : 350) and an antiDSG3 mouse monoclonal primary antibody (clone 7B9, Abcam; dilution 1 : 150).

Techniques: Expressing

Journal: British journal of cancer

Article Title: Desmogleins as prognostic biomarkers in resected pancreatic ductal adenocarcinoma.

doi: 10.1038/bjc.2015.362

Figure Lengend Snippet: Figure 4. DSG2 and DSG3 expression correlate with inferior survival of PDAC patients. Univariate analysis (Kaplan–Meier curve and log-rank test) in a TCGA RNA-Seq data set of PDAC tissue samples examining (A) DSG2 and (B) DSG3 expression levels as dichotomous variable after defining a cutoff via ROC analysis. Crossed lines indicate censored cases.

Article Snippet: Immunohistochemical staining of 4 mm sections was done strictly following the manufacturer’s instructions on a Ventana Benchmark XT autostainer (Ventana Medical Systems, Oro Valley, AZ, USA) using an anti-DSG1 rabbit monoclonal primary antibody (clone EPR6766(B), Abcam, Cambridge, UK; dilution 1 : 400), an anti-DSG2 rabbit polyclonal primary antibody (Atlas Antibodies, Stockholm, Sweden; dilution 1 : 350) and an antiDSG3 mouse monoclonal primary antibody (clone 7B9, Abcam; dilution 1 : 150).

Techniques: Expressing, RNA Sequencing

Journal: PLoS ONE

Article Title: In Vitro Functional Analyses of Arrhythmogenic Right Ventricular Cardiomyopathy-Associated Desmoglein-2-Missense Variations

doi: 10.1371/journal.pone.0047097

Figure Lengend Snippet: Investigated DSG2-variants.

Article Snippet: For Western blotting a murine IgG1 antibody against the extracellular domain of DSG2 clone 10G11 (anti-DSG2-10G11; BM5016, Acris Antibodies GmbH), a murine IgG1 antibody clone DG3.10 against the intracellular domain of DSG1+2 (anti-DSG1+2-DG3.10; BM370, Acris Antibodies GmbH) and a rabbit polyclonal antibody to PDI IgG (anti-PDI; ab31811, Abcam) as an endoplasmatic reticulum marker were used.

Techniques: Variant Assay

Journal: PLoS ONE

Article Title: In Vitro Functional Analyses of Arrhythmogenic Right Ventricular Cardiomyopathy-Associated Desmoglein-2-Missense Variations

doi: 10.1371/journal.pone.0047097

Figure Lengend Snippet: Schematic view of the rECD with analysed ARVC-associated variations. The dotted line shows the predicted PC cleavage site. SS = signal sequence, Pro = prodomain, EC1-EC4 = DSG2 extracellular cadherin subdomains 1-4. B Recombinantly expressed proteins were identified as DSG2-ECD with anti-DSG2-10G11 by Western blot analysis. The calculated apparent molecular weights were 67.5±1.5, 72.5±3.5, 70.0±3.0, 70.0±3.0, 70.5±2.5, and 69.0±4.0 (mean±SEM; n = 2) for the proteins in the traces in 1, 2, 3, 4, 5 and 6, respectively. C Coomassie-R-250 staining revealed the purity of the proteins. 1 = rECD-wt, 2-6 = rECDs as labelled in A .

Article Snippet: For Western blotting a murine IgG1 antibody against the extracellular domain of DSG2 clone 10G11 (anti-DSG2-10G11; BM5016, Acris Antibodies GmbH), a murine IgG1 antibody clone DG3.10 against the intracellular domain of DSG1+2 (anti-DSG1+2-DG3.10; BM370, Acris Antibodies GmbH) and a rabbit polyclonal antibody to PDI IgG (anti-PDI; ab31811, Abcam) as an endoplasmatic reticulum marker were used.

Techniques: Sequencing, Western Blot, Staining

Journal: PLoS ONE

Article Title: In Vitro Functional Analyses of Arrhythmogenic Right Ventricular Cardiomyopathy-Associated Desmoglein-2-Missense Variations

doi: 10.1371/journal.pone.0047097

Figure Lengend Snippet: A+B Flow cytometry-based assay for the binding of 0.8 µM rECD-wt or -variants to HT1080. A Representative histograms of FITC- fluorescence for binding of rECD-wt- and rECD-R46Q (as indicated). Bound rECD was detected with anti-HisFITC. As a negative control, HT1080 cells were incubated with only anti-HisFITC (grey filled area). B Column plots representing the ratio of rECD-binding related to the negative control (ratio rECD-bound ) as detected by flow cytometry. Ratios rECD-bound are indicated as mean± SEM of 7 independent measurements for rECD-variants and 9 independent measurements for rECD-wt with rECDs from at least 3 different purifications. Statistical analysis was performed by one-way ANOVA with Dunnett’s posttest using rECD-wt as a control (GraphPad Prism 5.01). rECD-R46Q-binding to HT1080 is increased 1.8-fold as compared to rECD-wt. Other ARVC-associated variants have no influence on rECD-binding to HT1080. C Representative Western blot (with anti-DSG2-10G11) of rECDs crosslinked in a 5 mM CaCl 2 containing buffer with BS 3 (+) or of controls (-) reveals that rECD wild-type and variants exist in solution as monomers (m), dimers (d), and oligomers (o).

Article Snippet: For Western blotting a murine IgG1 antibody against the extracellular domain of DSG2 clone 10G11 (anti-DSG2-10G11; BM5016, Acris Antibodies GmbH), a murine IgG1 antibody clone DG3.10 against the intracellular domain of DSG1+2 (anti-DSG1+2-DG3.10; BM370, Acris Antibodies GmbH) and a rabbit polyclonal antibody to PDI IgG (anti-PDI; ab31811, Abcam) as an endoplasmatic reticulum marker were used.

Techniques: Flow Cytometry, Binding Assay, Fluorescence, Negative Control, Incubation, Western Blot

Journal: PLoS ONE

Article Title: In Vitro Functional Analyses of Arrhythmogenic Right Ventricular Cardiomyopathy-Associated Desmoglein-2-Missense Variations

doi: 10.1371/journal.pone.0047097

Figure Lengend Snippet: DSC2b-HT1080 cells were transfected with full-length(fl)-DSG2-pEYFP; live cells were analysed with a fluorescence miscroscope one day after transfection. R46Q, D154E, D187G, K294E and V392I indicate the sequence variant in fl-DSG2-EYFP, wt fl-DSG2-wt-pEYFP, and C the LFA mock transfected control. Chimeric DSG2-proteins localised preferentially to the cell borders. ARVC-associated variations had no detectable influence on the localisation of fl-DSG2-EYFP in DSC2b-HT1080. Images were acquired through YFP and phase-contrast filters. Scale (red bar) = 10 µm.

Article Snippet: For Western blotting a murine IgG1 antibody against the extracellular domain of DSG2 clone 10G11 (anti-DSG2-10G11; BM5016, Acris Antibodies GmbH), a murine IgG1 antibody clone DG3.10 against the intracellular domain of DSG1+2 (anti-DSG1+2-DG3.10; BM370, Acris Antibodies GmbH) and a rabbit polyclonal antibody to PDI IgG (anti-PDI; ab31811, Abcam) as an endoplasmatic reticulum marker were used.

Techniques: Transfection, Fluorescence, Sequencing, Variant Assay