Journal: Scientific Reports

Article Title: Usp16 modulates Wnt signaling in primary tissues through Cdkn2a regulation

doi: 10.1038/s41598-018-34562-w

Figure Lengend Snippet: Cdkn2a mediates Usp16 response to Wnt3a. ( a ) Basal cell expansion is observed in the mammary gland of Cdkn2a +/− mice. A Graph quantifying the basal/luminal ratio is shown. Each dot represents an individual mouse. ( b ) Cdkn2a +/− sorted basal cells show an increased induction of Axin2 mRNA levels after 16 hours of Wnt3A stimulation (50 ng/ml). Two independent experiments were performed for a total of 4 animals per group. ( c ) Terminal Tip Fibroblasts (TTFs) derived from Cdkn2a +/− animals show an increase response to two different doses of Wnt3a (20 ng/ml and 50 ng/ml). The graph shows activation of a Top/Flash reporter for Wnt activity normalized by Renilla expression, co-transfected along with the reporter. ( d ) TTFs derived from Cdkn2a −/− animals and transfected with a vector expressing a Usp16 transgene or a control vector were analyzed for their ability to activate the Top-flash reporter. No differences were observed with two different doses of Wnt3a (20 ng/ml and 100 ng/ml). ( e ) MEFs cells were transiently transfected with two individual siRNAs targeting Bmi1. Activation of 6KD Wnt reporter is shown, normalized by Renilla expression. Cells were treated with 100 ng/ml of Wnt3a for 16 hours. ( f ) Two different human foreskin cultures were transiently transfected with siRNA targeting USP16, p16 Ink4a or a scramble sequence. USP16 and p16 Ink4a downregulation promotes the induction of AXIN2 mRNA expression 24 hours after treatment with 100 ng/ml of Wnt3a.

Article Snippet: Bmi1 , Hs00995519_g1 , Mm03053308_g1.

Techniques: Derivative Assay, Activation Assay, Activity Assay, Expressing, Transfection, Plasmid Preparation, Control, Sequencing

Journal: Scientific Reports

Article Title: Usp16 modulates Wnt signaling in primary tissues through Cdkn2a regulation

doi: 10.1038/s41598-018-34562-w

Figure Lengend Snippet: General mechanism of Wnt induction. ( a ) Proposed mechanism of interaction between Usp16 and the Wnt pathway. Bmi-1 and Usp16 control the level of ubiquitination of histone H2A. The balance between these two antagonistic proteins determines the levels of activation of Cdkn2a . Cdkn2a acts as a negative regulator of the Wnt pathway. Other mechanisms linking Bmi1 and Usp16 to Wnt activation are also possible. ( b ) During aging, increased tissue senescence and increased expression levels of p16 INK4a have been observed. At the same time, there is a concurrent decline in stem cell self renewal and Wnt activation. These events are correlated to a change in balance between Bmi1 - promoting self-renewal and Wnt activation - and Usp16 - promoting cellular senescence and Cdkn2a expression.

Article Snippet: Bmi1 , Hs00995519_g1 , Mm03053308_g1.

Techniques: Control, Ubiquitin Proteomics, Activation Assay, Expressing

Journal: Scientific Reports

Article Title: Usp16 modulates Wnt signaling in primary tissues through Cdkn2a regulation

doi: 10.1038/s41598-018-34562-w

Figure Lengend Snippet: List of Taqman assays (mouse and human-specific) used for this study. Probes were purchased from Applied Biosystem.

Article Snippet: Bmi1 , Hs00995519_g1 , Mm03053308_g1.

Techniques:

Journal: Scientific Reports

Article Title: Usp16 modulates Wnt signaling in primary tissues through Cdkn2a regulation

doi: 10.1038/s41598-018-34562-w

Figure Lengend Snippet: List of siRNA (mouse and human-specific) used for this study. siRNA were purchased from Dharmacon.

Article Snippet: Bmi1 , Hs00995519_g1 , Mm03053308_g1.

Techniques:

Journal: Journal of Cancer

Article Title: Investigating the Association between COMMD3 Expression and the Prognosis of Hepatocellular Carcinoma

doi: 10.7150/jca.62454

Figure Lengend Snippet: Primers used for QRT-PCR

Article Snippet: Anti-COMMD3 polyclonal antibodies and anti-CD34 monoclonal antibodies were purchased from Bioss (Boston, USA).

Techniques:

Journal: Journal of Cancer

Article Title: Investigating the Association between COMMD3 Expression and the Prognosis of Hepatocellular Carcinoma

doi: 10.7150/jca.62454

Figure Lengend Snippet: The COMMD3 expression in HCC tissue vs normal liver tissue, or HCC cells lines vs normal liver cell line. (A) COMMD3 mRNA levels in HCC tissue and normal liver tissue from TCGA. (B) COMMD3 mRNA levels in HCC tissue and normal liver tissue from Oncomine. (C) COMMD3 protein levels in HCC tissue and normal liver tissue from The Human Protein Atlas. (D) COMMD3 mRNA levels from HCC cells lines and normal liver cell line. (E) COMMD3 protein levels from HCC cells lines and normal liver cell line.

Article Snippet: Anti-COMMD3 polyclonal antibodies and anti-CD34 monoclonal antibodies were purchased from Bioss (Boston, USA).

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Investigating the Association between COMMD3 Expression and the Prognosis of Hepatocellular Carcinoma

doi: 10.7150/jca.62454

Figure Lengend Snippet: Kaplan-Meier curves of HCC patients grouped by COMMD3 mRNA levels. (A) High expression level of COMMD3 mRNA was associated with poor OS in HCC patients. (B) High expression level of COMMD3 mRNA was associated with poor DFS in HCC patients.

Article Snippet: Anti-COMMD3 polyclonal antibodies and anti-CD34 monoclonal antibodies were purchased from Bioss (Boston, USA).

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Investigating the Association between COMMD3 Expression and the Prognosis of Hepatocellular Carcinoma

doi: 10.7150/jca.62454

Figure Lengend Snippet: Correlation between COMMD3 expression level and clinical-pathological variables in HCC patients

Article Snippet: Anti-COMMD3 polyclonal antibodies and anti-CD34 monoclonal antibodies were purchased from Bioss (Boston, USA).

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Investigating the Association between COMMD3 Expression and the Prognosis of Hepatocellular Carcinoma

doi: 10.7150/jca.62454

Figure Lengend Snippet: Cox proportional-hazard regression analysis for OS in HCC patients

Article Snippet: Anti-COMMD3 polyclonal antibodies and anti-CD34 monoclonal antibodies were purchased from Bioss (Boston, USA).

Techniques:

Journal: Journal of Cancer

Article Title: Investigating the Association between COMMD3 Expression and the Prognosis of Hepatocellular Carcinoma

doi: 10.7150/jca.62454

Figure Lengend Snippet: Cox proportional-hazard regression analysis for DFS in HCC patients

Article Snippet: Anti-COMMD3 polyclonal antibodies and anti-CD34 monoclonal antibodies were purchased from Bioss (Boston, USA).

Techniques:

Journal: Journal of Cancer

Article Title: Investigating the Association between COMMD3 Expression and the Prognosis of Hepatocellular Carcinoma

doi: 10.7150/jca.62454

Figure Lengend Snippet: Establishment of the stably transformed SK-Hep1 and Hep-3B cell lines with COMMD3 shRNA. (A) COMMD3 mRNA levels in SK-Hep1 and Hep-3B cell lines transfected with COMMD3 shRNA. (B) The levels of COMMD3 proteins in SK-Hep1 and Hep-3B cell lines transfected with COMMD3 shRNA. (C) The relative expression levels of COMMD3 proteins in the SK-Hep1 and Hep-3B cell lines transfected with COMMD3 shRNA were quantified using ImageJ software. Data were plotted with GraphPad Prism 7.0 software. * P <0.05, ** P <0.01 and *** P <0.001 compared to the control group.

Article Snippet: Anti-COMMD3 polyclonal antibodies and anti-CD34 monoclonal antibodies were purchased from Bioss (Boston, USA).

Techniques: Stable Transfection, Transformation Assay, shRNA, Transfection, Expressing, Software

Journal: Journal of Cancer

Article Title: Investigating the Association between COMMD3 Expression and the Prognosis of Hepatocellular Carcinoma

doi: 10.7150/jca.62454

Figure Lengend Snippet: The effects of COMMD3 knockdown on the mRNA and protein levels of HIF1α and VEGF in HCC cells. (A) The effects of COMMD3 knockdown on VEGF mRNA levels in SK-Hep1 and Hep-3B cell lines. (B) The effects of the COMMD3 knockdown on HIF1α mRNA levels in SK-Hep1 and Hep-3B cell lines. (C) The effects of COMMD3 knockdown on protein levels of HIF1α and VEGF protein in SK-Hep1 and Hep-3B cell lines. (D, E) The relative expression levels of HIF1α and VEGF proteins in SK-Hep1 and Hep-3B cell lines with COMMD3 shRNA were quantified using ImageJ software. Data were plotted using GraphPad Prism 7.0 software. * P <0.05, ** P <0.01 and *** P <0.001compared to the control group.

Article Snippet: Anti-COMMD3 polyclonal antibodies and anti-CD34 monoclonal antibodies were purchased from Bioss (Boston, USA).

Techniques: Expressing, shRNA, Software

Journal: Journal of Cancer

Article Title: Investigating the Association between COMMD3 Expression and the Prognosis of Hepatocellular Carcinoma

doi: 10.7150/jca.62454

Figure Lengend Snippet: The inhibitory effects of COMMD3 knockdown on the migration, invasion and angiogenic ability of HCC cells. (A) The wound healing assay used for testing cell migration ability; Scale bar, 20 µm in A. (B) The histogram represents the migration distance relative to the control at 24 h after scratching the cell layer. (C) The transwell assay used for testing cell invasion. Scale bar, 10μm in C. (D) The histogram represents the average number of invasive cells relative to the control. (E) The tube formation assay used for testing HCC cell-induced angiogenesis in HUVECs. Scale bar, 10μm in E. (F) The area covered by the tube network was quantified using the Image-Pro Plus software. Data were plotted with the GraphPad Prism 7.0 software. * P <0.05, ** P <0.01 and *** P <0.001compared to the control group. 2.5 times magnification in the right side of all original figures.

Article Snippet: Anti-COMMD3 polyclonal antibodies and anti-CD34 monoclonal antibodies were purchased from Bioss (Boston, USA).

Techniques: Migration, Wound Healing Assay, Transwell Assay, Tube Formation Assay, Software

Journal: BMC Cancer

Article Title: Transcriptional analysis of the expression, prognostic value and immune infiltration activities of the COMMD protein family in hepatocellular carcinoma

doi: 10.1186/s12885-021-08699-3

Figure Lengend Snippet: Univariate and multivariate analysis of overall survival in 344 HCC patients

Article Snippet: Lipofectamine RNAiMAX (ThermoFisher, USA) was used to transfect Negative Control (NC) and COMMD3 siRNAs (Ribobio, China) into HCC cells as the manufacturer’s protocol suggested.

Techniques:

Journal: BMC Cancer

Article Title: Transcriptional analysis of the expression, prognostic value and immune infiltration activities of the COMMD protein family in hepatocellular carcinoma

doi: 10.1186/s12885-021-08699-3

Figure Lengend Snippet: Univariate and multivariate of overall survival in 130 patients with HCC of grade III and IV

Article Snippet: Lipofectamine RNAiMAX (ThermoFisher, USA) was used to transfect Negative Control (NC) and COMMD3 siRNAs (Ribobio, China) into HCC cells as the manufacturer’s protocol suggested.

Techniques:

Journal: BMC Cancer

Article Title: Transcriptional analysis of the expression, prognostic value and immune infiltration activities of the COMMD protein family in hepatocellular carcinoma

doi: 10.1186/s12885-021-08699-3

Figure Lengend Snippet: The expression and prognostic value of COMMD3 was validated in GSE14520 and HCC patients. A COMMD3 expression in GSE14520 datasets between one thirds of the highest expression and two thirds of the lowest expression. B K-M OS curves in GSE14520 datasets stratified by COMMD3 expression. C COMMD3 expression in normal and tumor tissue of 80 HCC patients. D K–M OS curves in 80 HCC patients stratified by COMMD3 expression. E CCK-8 cell proliferation assay results in Hep3B and HepG2 cells transfected with si-COMMD3, or negative control (NC). F EdU cell proliferation assay results in Hep3B and HepG2 cells transfected with si-COMMD3, or NC. * p < 0.05, *** p < 0.001

Article Snippet: Lipofectamine RNAiMAX (ThermoFisher, USA) was used to transfect Negative Control (NC) and COMMD3 siRNAs (Ribobio, China) into HCC cells as the manufacturer’s protocol suggested.

Techniques: Expressing, CCK-8 Assay, Proliferation Assay, Transfection, Negative Control