mouse monoclonal dsc3  (Novus Biologicals)

Journal: Investigative Ophthalmology & Visual Science

Article Title: Calcium-Dependent Regulation of Desmosomal Proteins in Human Meibomian Gland Epithelial Cells

doi: 10.1167/iovs.66.6.49

Figure Lengend Snippet: Immunohistochemical detection of DSC in the meibomian glands of C57BL/6 mice. ( A ) Acinus (ac) with basal cells (b), differentiating meibocytes (d), and mature meibocytes (m). The immune response is slightly stronger in the peripheral areas of the acinus than in the central areas. The cell borders between individual meibocytes show a stronger immune response and indicate the presence of DSC in the cell membrane. ( B ) Transition of the disintegration zone of an acinus (ac) into the connecting duct (cd). DSC (red reaction product) can be seen in the acini as well as in the epithelia of the connecting and central excretory ducts. Especially the apical layers of the epithelia in particular show a strong immunoreaction. Parts of the ductal system are filled with degenerated cells and meibum in which DSC can also be detected. ( C ) Detection of DSC in the epidermis (epi) of the eyelid serves as a positive control. ( D ) The negative control shows no immunoreaction. The figure shows representative results of three independent experiments ( n = 3). DSC, desmocollin.

Article Snippet: DSC , Acris Antibodies , SP1050 , IHC (1:50); IF (1:50); WB (1:250).

Techniques: Immunohistochemical staining, Membrane, Positive Control, Negative Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: SOX30 is a key regulator of desmosomal gene suppressing tumor growth and metastasis in lung adenocarcinoma

doi: 10.1186/s13046-018-0778-3

Figure Lengend Snippet: Positive percentage of desmosomal gene expression between ADC and SCC

Article Snippet: Following blocking with 1% bovine serum albumin for 30 min, rabbit polyclonal antibody SOX30 (1:100; Santa Cruz Biotechnology; sc-20,104), mouse monoclonal antibody DSP (1:100; Santa Cruz Biotechnology; sc-365,981), mouse monoclonal antibody JUP (1:100; Santa Cruz Biotechnology; sc-33,634) and mouse monoclonal antibody DSC3 (1:100; Santa Cruz Biotechnology; sc-81,806) were incubated overnight at 4 °C.

Techniques: Gene Expression

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: SOX30 is a key regulator of desmosomal gene suppressing tumor growth and metastasis in lung adenocarcinoma

doi: 10.1186/s13046-018-0778-3

Figure Lengend Snippet: SOX30 is positive correlated with desmosomal gene expression in ADC, but not in SCC. a Heatmaps for correlations between SOX30 and desmosomal genes in the TCGA lung adenocarcinoma RNAseq (IlluminaHiSeq; n = 571) data set. b Heatmaps for correlations between SOX30 and desmosomal genes in the TCGA lung squamous carcinoma RNAseq (IlluminaHiSeq; n = 553) data set. Correlation coefficient R and P -values were calculated by a Spearman correlation analysis. c qRT-PCR analysis of desmosomal gene expression in A549 and LTEP-a-2 cells transiently transfected with the vector control or SOX30 expression vector. d qRT-PCR analysis of desmosomal gene expression in H520 and H226 cells transiently transfected with the vector control or SOX30 expression vector. ACTIN was used as an internal control. e The protein levels of DSP, and JUP and DSC3 were monitored by WB after SOX30 overexpression in A549 and LTEP-a-2 cells. f The protein levels of DSP, and JUP and DSC3 were monitored by WB after SOX30 overexpression in H520 and H226 cells. g The protein levels of SOX30, DSP, JUP and DSC3 were further monitored by IHC in human ADC tissues. h The protein levels of SOX30, DSP, JUP and DSC3 were further monitored by IHC in human SCC tissues. Scale bar represents 50 mm

Article Snippet: Following blocking with 1% bovine serum albumin for 30 min, rabbit polyclonal antibody SOX30 (1:100; Santa Cruz Biotechnology; sc-20,104), mouse monoclonal antibody DSP (1:100; Santa Cruz Biotechnology; sc-365,981), mouse monoclonal antibody JUP (1:100; Santa Cruz Biotechnology; sc-33,634) and mouse monoclonal antibody DSC3 (1:100; Santa Cruz Biotechnology; sc-81,806) were incubated overnight at 4 °C.

Techniques: Gene Expression, Quantitative RT-PCR, Transfection, Plasmid Preparation, Control, Expressing, Over Expression

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: SOX30 is a key regulator of desmosomal gene suppressing tumor growth and metastasis in lung adenocarcinoma

doi: 10.1186/s13046-018-0778-3

Figure Lengend Snippet: SOX30 directly binding to promoters of DSP, JUP and DSC3. a–c The ACAAT motif in promoter region of DSP, JUP and DSC3 were underlined. TSS, transcription start site. d ChIP-PCR was performed to identify DSP, JUP and DSC3 are direct binding targets of SOX30

Article Snippet: Following blocking with 1% bovine serum albumin for 30 min, rabbit polyclonal antibody SOX30 (1:100; Santa Cruz Biotechnology; sc-20,104), mouse monoclonal antibody DSP (1:100; Santa Cruz Biotechnology; sc-365,981), mouse monoclonal antibody JUP (1:100; Santa Cruz Biotechnology; sc-33,634) and mouse monoclonal antibody DSC3 (1:100; Santa Cruz Biotechnology; sc-81,806) were incubated overnight at 4 °C.

Techniques: Binding Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: SOX30 is a key regulator of desmosomal gene suppressing tumor growth and metastasis in lung adenocarcinoma

doi: 10.1186/s13046-018-0778-3

Figure Lengend Snippet: Desmosomal gene silencing diminished the effect of SOX30 overexpression on proliferation, migration and invasion of A549 and LTEP-a-2 cells. a SOX30, DSP, JUP and DSC3 expression were confirmed by WB in A549 and LTEP-a-2 cells. b MTS assays were performed to analyze cell proliferation of A549 and LTEP-a-2 cells cotransfected with desmosomal gene miRNA and SOX30 expression vector. * P < 0.05; ** P < 0.01. c The effect of increasing SOX30 expression with decreasing desmosomal gene expression on cell growth was further confirmed by colony formation assay in A549 and LTEP-a-2 cells. d Assessment of the effects of the expression of SOX30 and desmosomal gene, respectively, on cell migration and invasion in A549 and LTEP-a-2 cells

Article Snippet: Following blocking with 1% bovine serum albumin for 30 min, rabbit polyclonal antibody SOX30 (1:100; Santa Cruz Biotechnology; sc-20,104), mouse monoclonal antibody DSP (1:100; Santa Cruz Biotechnology; sc-365,981), mouse monoclonal antibody JUP (1:100; Santa Cruz Biotechnology; sc-33,634) and mouse monoclonal antibody DSC3 (1:100; Santa Cruz Biotechnology; sc-81,806) were incubated overnight at 4 °C.

Techniques: Over Expression, Migration, Expressing, Plasmid Preparation, Gene Expression, Colony Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: SOX30 is a key regulator of desmosomal gene suppressing tumor growth and metastasis in lung adenocarcinoma

doi: 10.1186/s13046-018-0778-3

Figure Lengend Snippet: Knockdown of desmosomal gene expression attenuates the effects of SOX30 on impeding tumor formation and metastasis in vivo. a – c Evaluation of tumorigenesis in nude mice subcutaneously injected with (A549-Vector), (A549-SOX30(+)), (A549-SOX30(+)/DSP(−)), (A549-SOX30(+)/JUP(−)) and (A549-SOX30(+)/DSC3(−)). To assess the effects of desmosomal gene on subcutaneous tumor growth, tumor volumes were recorded on the indicated days, and pictures of solid tumor tissues were taken after 42 days. Tumor weights in the indicated groups were determined. Error bars indicate s.d. ( n = 4). d Liver metastases were observed by H&E staining in the indicated groups. Arrows indicate the metastatic loci. e Liver metastasis was further quantified using RT-qPCR. Human-specific β2-MG levels were used to quantify metastatic human cancer cells with the mouse-specific β2-MG level as an internal control. Error bars indicate s.d. (n = 4).* P < 0.05; ** P < 0.01

Article Snippet: Following blocking with 1% bovine serum albumin for 30 min, rabbit polyclonal antibody SOX30 (1:100; Santa Cruz Biotechnology; sc-20,104), mouse monoclonal antibody DSP (1:100; Santa Cruz Biotechnology; sc-365,981), mouse monoclonal antibody JUP (1:100; Santa Cruz Biotechnology; sc-33,634) and mouse monoclonal antibody DSC3 (1:100; Santa Cruz Biotechnology; sc-81,806) were incubated overnight at 4 °C.

Techniques: Knockdown, Gene Expression, In Vivo, Injection, Plasmid Preparation, Staining, Quantitative RT-PCR, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: SOX30 is a key regulator of desmosomal gene suppressing tumor growth and metastasis in lung adenocarcinoma

doi: 10.1186/s13046-018-0778-3

Figure Lengend Snippet: SOX30 inhibits Wnt and ERK signaling by accelerating the expression of desmosomal gene. a The protein expression of SOX30, DSP, JUP, CCND1 and MMP7 were detected in A549-SOX30 cells and subcutaneous tumors of mice. b A549 cells with SOX30 or vector control stable expression were co-transfected with luciferase reporter constructs (TOPFlash or FOPFlash, respectively) and Renilla luciferase constructs (normalizing transfection control). The reporter activity was measured 24 h after transfection. Error bars represent the s.d. of three independent experiments. ** SOX30 vs vector control, Student’s t-test, P < 0.01. c The protein expression of SOX30, DSC3, ERK1/2 and p-ERK1/2 were detected in A549-SOX30 cells and subcutaneous tumors of mice

Article Snippet: Following blocking with 1% bovine serum albumin for 30 min, rabbit polyclonal antibody SOX30 (1:100; Santa Cruz Biotechnology; sc-20,104), mouse monoclonal antibody DSP (1:100; Santa Cruz Biotechnology; sc-365,981), mouse monoclonal antibody JUP (1:100; Santa Cruz Biotechnology; sc-33,634) and mouse monoclonal antibody DSC3 (1:100; Santa Cruz Biotechnology; sc-81,806) were incubated overnight at 4 °C.

Techniques: Expressing, Plasmid Preparation, Control, Transfection, Luciferase, Construct, Activity Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: SOX30 is a key regulator of desmosomal gene suppressing tumor growth and metastasis in lung adenocarcinoma

doi: 10.1186/s13046-018-0778-3

Figure Lengend Snippet: Inhibition of SOX30 promotes tumorigenesis of lung cancer with urethane treatment. a The mRNA expression of DSP, JUP and DSC3 in lung tissues of SOX30-knockout mice were measured by qRT-PCR. b The protein expression of DSP, JUP and DSC3 in lung tissues of SOX30-knockout mice were detected by WB. c Lung tissues were processed and stained with H&E for detection of tumor foci. d Tumor extracts were analyzed by WB. ACTIN was used as a loading control. e Schematic diagram of the mechanisms of SOX30 mediated suppression of ADC cell proliferation and metastasis based on our study

Article Snippet: Following blocking with 1% bovine serum albumin for 30 min, rabbit polyclonal antibody SOX30 (1:100; Santa Cruz Biotechnology; sc-20,104), mouse monoclonal antibody DSP (1:100; Santa Cruz Biotechnology; sc-365,981), mouse monoclonal antibody JUP (1:100; Santa Cruz Biotechnology; sc-33,634) and mouse monoclonal antibody DSC3 (1:100; Santa Cruz Biotechnology; sc-81,806) were incubated overnight at 4 °C.

Techniques: Inhibition, Expressing, Knock-Out, Quantitative RT-PCR, Staining, Control

Journal: Traffic (Copenhagen, Denmark)

Article Title: ER-to-Golgi Blockade of Nascent Desmosomal Cadherins in SERCA2-inhibited Keratinocytes: Implications for Darier’s Disease

doi: 10.1111/tra.12470

Figure Lengend Snippet: (A) Skin tissues derived from a normal subject (a) and DD patients (b–d) were stained with Dsc3. (B) Skin tissues were dual labeled with Dsc3 (red) and ER calnexin (green) and examined in a confocal microscope. Colocalization of Dsc3 and calnexin (yellow signal) is revealed by merged images. Scale bar = 10 μm.

Article Snippet: Following deparaffinization, tissues sections were steamed with antigen retrieval buffer (BioGenex, Citra Plus, pH6.0) and stained with mouse monoclonal antibodies to Dsc3 (U114, GeneTex) and/or rabbit anti-calnexin.

Techniques: Derivative Assay, Staining, Labeling, Microscopy

Journal: Oncology Letters

Article Title: Particular gene upregulation and p53 heterogeneous expression in TP53 -mutated maxillary carcinoma

doi: 10.3892/ol.2017.6751

Figure Lengend Snippet: Primer sequences used for quantitative polymerase chain reaction analysis in this study.

Article Snippet: The primary antibodies used were rabbit polyclonal anti-human CSTA IgG (0.1 µg/ml, HPA001031; Atlas Antibodies AB, Stockholm, Sweden), mouse monoclonal anti-human 14-3-3 sigma IgG1 clone 1.N.6 (1 µg/ml, GTX14123; GeneTex, Irvine, CA, USA), mouse monoclonal anti-human desmocollin 3 IgG1 clone Dsc3-U114 (0.05 µg/ml, 61093; Progen Biotech, Heidelberg, Germany), and mouse monoclonal anti-human p53 IgG2b clone DO-7 (0.69 µg/ml, M7001; Dako, Glostrup, Denmark).

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: Oncology Letters

Article Title: Particular gene upregulation and p53 heterogeneous expression in TP53 -mutated maxillary carcinoma

doi: 10.3892/ol.2017.6751

Figure Lengend Snippet: Twenty-one validated genes with differential expression in TP53 mutated versus non-mutated tumors.

Article Snippet: The primary antibodies used were rabbit polyclonal anti-human CSTA IgG (0.1 µg/ml, HPA001031; Atlas Antibodies AB, Stockholm, Sweden), mouse monoclonal anti-human 14-3-3 sigma IgG1 clone 1.N.6 (1 µg/ml, GTX14123; GeneTex, Irvine, CA, USA), mouse monoclonal anti-human desmocollin 3 IgG1 clone Dsc3-U114 (0.05 µg/ml, 61093; Progen Biotech, Heidelberg, Germany), and mouse monoclonal anti-human p53 IgG2b clone DO-7 (0.69 µg/ml, M7001; Dako, Glostrup, Denmark).

Techniques: Expressing, Protease Inhibitor, Coagulation, Binding Assay

Journal: Oncology Letters

Article Title: Particular gene upregulation and p53 heterogeneous expression in TP53 -mutated maxillary carcinoma

doi: 10.3892/ol.2017.6751

Figure Lengend Snippet: Representative IHC analysis of CSTA, SFN, DSC3, and p53 expression in maxillary squamous cell carcinoma with and without Tp53 mutation. Normal, normal paranasal sinus mucosa; TP53 mutation (−), case 1T; TP53 mutation (+), case 8T (CSTA, DSC3) and M9 (SFN). Bar, 100 µm.

Article Snippet: The primary antibodies used were rabbit polyclonal anti-human CSTA IgG (0.1 µg/ml, HPA001031; Atlas Antibodies AB, Stockholm, Sweden), mouse monoclonal anti-human 14-3-3 sigma IgG1 clone 1.N.6 (1 µg/ml, GTX14123; GeneTex, Irvine, CA, USA), mouse monoclonal anti-human desmocollin 3 IgG1 clone Dsc3-U114 (0.05 µg/ml, 61093; Progen Biotech, Heidelberg, Germany), and mouse monoclonal anti-human p53 IgG2b clone DO-7 (0.69 µg/ml, M7001; Dako, Glostrup, Denmark).

Techniques: Expressing, Mutagenesis