Journal: Viruses

Article Title: HBV Infection Drives PSMB5-Dependent Proteasomal Activation in Humanized Mice and HBV-Associated HCC

doi: 10.3390/v17111454

Figure Lengend Snippet: HBV infection enhances PSMB5 expression in liver-humanized mice. ( A ) 10-year survival analysis of low (bottom 75% of TPM) and high (top 25% of TPM) PSMA1 expressing HCC patients using GEPIA: a web server for cancer and normal gene expression profiling and interactive analyses . HBV infection increases PSMB5 gene expression levels of liver humanized mice livers. ( B ) Schematic overview of HBV infection in uPA-NOG liver-humanized mice. ( C ) Human albumin levels in mouse serum, 8 weeks post-transplantation ( D ) H&E staining of mouse liver tissue. ( E ) Anti-human mitochondria staining indicating engrafted human hepatocytes after 12 weeks of infection. ( F ) Relative mRNA expression of 26S proteasome subunits in livers of HBV-infected vs. non-infected mice ( n = 4/group). TPM: Transcripts per million. Scale bar: 100 μm.

Article Snippet: The plasmid targeting the human PSMB5 gene (#117073) and the LentiCRISPRv2 plasmid with blasticidin selection (#83480) were obtained from Addgene (Watertown, MA, USA).

Techniques: Infection, Expressing, Gene Expression, Transplantation Assay, Staining

Journal: Viruses

Article Title: HBV Infection Drives PSMB5-Dependent Proteasomal Activation in Humanized Mice and HBV-Associated HCC

doi: 10.3390/v17111454

Figure Lengend Snippet: Functional consequences of PSMB5 knockout in human THP-1 cells. ( A ) Ten-year survival analysis of low (bottom 75% of TPM) and high (top 25% of TPM) PSMB5 expressing HCC patients using GEPIA: a web server for cancer and normal gene expression profiling and interactive analyses . ( B ) PSMB5KO cells accumulate ubiquitinated proteins due to decreased protein degradation. Western blot analysis showing accumulation of ubiquitinated proteins in PSMB5-deficient (KO) THP-1 cells compared with wild-type (WT) controls on day 7. ( C ) Flow cytometric analysis of MHC I surface expression in WT and PSMB5KO THP-1cells at days 7 and day 14 post-transduction. ( D ) Relative mRNA expression levels of antigen-processing pathway genes in WT ( n = 3) and PSMB5-KO ( n = 3) THP-1 cells. WT: wild-type control; KO: PSMB5 knockout. * p < 0.05.

Article Snippet: The plasmid targeting the human PSMB5 gene (#117073) and the LentiCRISPRv2 plasmid with blasticidin selection (#83480) were obtained from Addgene (Watertown, MA, USA).

Techniques: Functional Assay, Knock-Out, Expressing, Gene Expression, Western Blot, Transduction, Control

Journal: Viruses

Article Title: HBV Infection Drives PSMB5-Dependent Proteasomal Activation in Humanized Mice and HBV-Associated HCC

doi: 10.3390/v17111454

Figure Lengend Snippet: Elevated β5 subunit expression and proteasome activity in HBV-infected human tissues. ( A ) Differential expression of PSMB5 gene in tumor and corresponding normal tissue from patients with HCC using GEPIA . HBV infected patients have higher levels of β5 protein expression and proteasome activity levels. Liver and serum samples of HBV, HCC and HBV-induced HCC patients were collected; ( B ) proteasome subunit β5 protein expression and ( C ) proteasome activity were analyzed with ELISA methods. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The plasmid targeting the human PSMB5 gene (#117073) and the LentiCRISPRv2 plasmid with blasticidin selection (#83480) were obtained from Addgene (Watertown, MA, USA).

Techniques: Expressing, Activity Assay, Infection, Quantitative Proteomics, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Rapid self-selecting and clone-free integration of transgenes into engineered CRISPR safe harbor locations in Caenorhabditis elegans

doi: 10.1101/2020.05.26.117390

Figure Lengend Snippet: (A) Schematic of landing pad insertion vector pMS74 and (B) corresponding color-coded sequence

Article Snippet: pMS4, pMS74, PMS79, pMS81, and pZCS16 are available from Addgene.

Techniques: Plasmid Preparation, Sequencing