Journal: Molecular Therapy. Nucleic Acids

Article Title: Unlocking the therapeutic potential of locked nucleic acids through lipid nanoparticle delivery

doi: 10.1016/j.omtn.2024.102224

Figure Lengend Snippet: Clinical trials of ASOs

Article Snippet: Santaris Pharma , SPC 2996 , intravenous infusion , B cell Lymphoma 2 (Bcl-2) , chronic lymphocytic leukemia , phase I/II , discontinued.

Techniques: Injection, Activity Assay, Emulsion, Reverse Transcription, Virus, Allele-specific Oligonucleotide

Journal: Cancers

Article Title: HPVE6-USP46 Mediated Cdt2 Stabilization Reduces Set8 Mediated H4K20-Methylation to Induce Gene Expression Changes

doi: 10.3390/cancers14010030

Figure Lengend Snippet: ( A ) Immunohistochemical staining of Cdt2, Set8, H4K20-me1 and H4K20-me3 cells in cervical cancer and adjacent normal tissues. ( B ) Summary of staining scores for Set8, H4K20-me1 and H4K20-me3 in normal and cancerous tissue. DCC 1–10 denotes individual patients for each normal and cancer pairs. ( C ) HeLa cells were transfected with control (Gl2) or indicated siRNAs and collected 48 h after transfection. The cell lysates were then immunoblotted with indicated antibodies. ( D ) HeLa cells were transfected twice with control (Gl2) or HPV-E6 siRNAs before and during the double thymidine block (DTB). Cells were collected at indicated time points after release from DTB and subjected to immunoblotting with indicated antibodies. ( E ) HeLa cells were transfected with GFP or HPV-16E6 expressing plasmids followed by double thymidine block (DTB). Cells were collected at indicated time points after DTB and subjected to immunoblotting with indicated antibodies.

Article Snippet: Antibodies used were Set8 (CST #2996), H4K20-me1(AbCam #ab9051) and H4K20-me3 (active motif #07-463).

Techniques: Immunohistochemical staining, Staining, Transfection, Control, Blocking Assay, Western Blot, Expressing

Journal: Cancers

Article Title: HPVE6-USP46 Mediated Cdt2 Stabilization Reduces Set8 Mediated H4K20-Methylation to Induce Gene Expression Changes

doi: 10.3390/cancers14010030

Figure Lengend Snippet: ( A ) Immunoblotting of indicated proteins after siE6 or siE6+siSet8. ( B ) Principal component analysis of the RNA seq results which show that the RNA seq from the knockdown conditions are reproducible and cluster as expected. ( C , D ) Volcano plots with Log2 fold change ( X -axis) vs. −Log10 p -value ( Y -axis) show that many genes are significantly up or down-regulated when E6 is knocked down ( C ) or when Set8 is co-knocked down with E6, relative to E6 alone ( D ). Red dots: genes with Log2FC > |0.5| and −Log10 adjusted p -value> 1.3. Genes with a −Log10 p -value > 15 were plotted at 15.

Article Snippet: Antibodies used were Set8 (CST #2996), H4K20-me1(AbCam #ab9051) and H4K20-me3 (active motif #07-463).

Techniques: Western Blot, RNA Sequencing, Knockdown

Journal: Cancers

Article Title: HPVE6-USP46 Mediated Cdt2 Stabilization Reduces Set8 Mediated H4K20-Methylation to Induce Gene Expression Changes

doi: 10.3390/cancers14010030

Figure Lengend Snippet: E6 induces EGFR expression via degradation of Set8 and controls EGF response genes. ( A ) EGFR expression in siE6 and siE6+siSet8 conditions from RNA sequencing. ( B ) qPCR validation of RNA sequencing results on ten genes in HeLa cells that were repressed by siE6 and where the repression was rescued by co-knockdown of Set8 * p < 0.05, ** p < 0.001, *** p < 0.0001. ( C ) H4K20me1 ChIP-seq reads in the EGFR gene from liver specific Set8 knockout mice (GSE97338) compared to that in WT mice. ( D ) ENCODE H4K20me1 ChIP-seq peaks in the EGFR gene from HeLa-S3 (HPV+) and keratinocytes. ( E ) and ( F ) EGF response gene set enrichment analysis for siE6+siSet8 relative to control (siGL2) and siE6 relative to control, respectively. ( G ) Heatmap of log2 fold change of highly expressed genes within the EGF response gene set from ( F ) and ( G ).

Article Snippet: Antibodies used were Set8 (CST #2996), H4K20-me1(AbCam #ab9051) and H4K20-me3 (active motif #07-463).

Techniques: Expressing, RNA Sequencing, Biomarker Discovery, Knockdown, ChIP-sequencing, Knock-Out, Control

Journal: Cancers

Article Title: HPVE6-USP46 Mediated Cdt2 Stabilization Reduces Set8 Mediated H4K20-Methylation to Induce Gene Expression Changes

doi: 10.3390/cancers14010030

Figure Lengend Snippet: Model summarizing the role of the E6-USP46 complex in HPV induced cancers: E6 from high-risk HPVs engage with and activate the deubiquitinase activity of USP46. The complex then interacts with and stabilizes Cdt2, which in turn degrades the H4K20 mono-methyl-transferase Set8 resulting in decreased levels of H4K20 mono, di and tri methylations at H4K20. Depending on the gene this results in up or down regulation of several genes (like EGFR).

Article Snippet: Antibodies used were Set8 (CST #2996), H4K20-me1(AbCam #ab9051) and H4K20-me3 (active motif #07-463).

Techniques: Activity Assay