hpv 16 e7 fragment  (Addgene inc)

Journal: bioRxiv

Article Title: Critical role of cell competition in gliomagenesis

doi: 10.64898/2026.01.15.699808

Figure Lengend Snippet: A. Table of patient samples used in snRNA-seq, with the status of a few gene alterations listed. B. Sample MRIs from one patient showing T1 contrast-enhanced and T2 weighted images. On the left is the preoperative study demonstrating location of contrast enhancing zone (yellow arrow) containing the presumed solid tumor and the broader, T2-hyperintense signal (red arrow) that containing the presumed infiltrating tumor. The middle image shows the pre-operative regions selected for biopsy and the right image shows the intraoperative neuronavigation biopsy acquisition. C. The workflow from sample biopsy, processing, snRNA-sequencing, and data analysis. D. Bar plots of cell type proportions in the infiltrating and solid tumor regions. Each bar represents the mean proportion of the selected cell type in the region and the positive and negative standard deviation are represented by the bracket. Each dot represents the cell type proportion of a single sample. E. Representative images of expanded Nf1- knockout OPCs (GFP + PDGFRα + ) generated by transduction of MSCV carrying Nf1 guide RNA/Cas9. WT OPCs (GFP - PDGFRα + ) were excluded from the expanded Nf1 -null OPC clones, suggesting OPC competition. Scale bar: 50 μm. F. Representative images of expanded Rb -inactivated OPCs (GFP + PDGFRα + ) upon transduction of MSCV carrying HPV-16 E7 protein that inactivates Rb function. Expanded GFP + Rb -inactivated OPCs (orange arrow) intermixed with WT OPCs (GFP - PDGFRα + , white arrow-head), suggesting the lack of OPC competition. Scale bar: 50 μm G. Representative images showing the expansion of OPCs with EGFR mutation ( EGFRvIII ). The expanded clone of EGFRvIII -expressing OPCs excluded WT OPCs (GFP - PDGFRα + ), suggesting OPC competition. Scale bar: 50 μm. H. Summary of the sufficiency and insufficiency in driving OPC competition of all examined gliomagenic mutations in this study.

Article Snippet: To inactivate Rb function, the HPV-16 E7 fragment was amplified from plasmid p1324 HPV-16 E7 (Addgene#8643) and cloned to replace Cas9 in MSCV vector, maintaining the in-frame fusion with T2A-EGFP for accurate reporting.

Techniques: Sequencing, Standard Deviation, Knock-Out, Generated, Transduction, Clone Assay, Mutagenesis, Expressing

Journal: bioRxiv

Article Title: Critical role of cell competition in gliomagenesis

doi: 10.64898/2026.01.15.699808

Figure Lengend Snippet: A. Table of patient samples used in snRNA-seq, with the status of a few gene alterations listed. B. Sample MRIs from one patient showing T1 contrast-enhanced and T2 weighted images. On the left is the preoperative study demonstrating location of contrast enhancing zone (yellow arrow) containing the presumed solid tumor and the broader, T2-hyperintense signal (red arrow) that containing the presumed infiltrating tumor. The middle image shows the pre-operative regions selected for biopsy and the right image shows the intraoperative neuronavigation biopsy acquisition. C. The workflow from sample biopsy, processing, snRNA-sequencing, and data analysis. D. Bar plots of cell type proportions in the infiltrating and solid tumor regions. Each bar represents the mean proportion of the selected cell type in the region and the positive and negative standard deviation are represented by the bracket. Each dot represents the cell type proportion of a single sample. E. Representative images of expanded Nf1- knockout OPCs (GFP + PDGFRα + ) generated by transduction of MSCV carrying Nf1 guide RNA/Cas9. WT OPCs (GFP - PDGFRα + ) were excluded from the expanded Nf1 -null OPC clones, suggesting OPC competition. Scale bar: 50 μm. F. Representative images of expanded Rb -inactivated OPCs (GFP + PDGFRα + ) upon transduction of MSCV carrying HPV-16 E7 protein that inactivates Rb function. Expanded GFP + Rb -inactivated OPCs (orange arrow) intermixed with WT OPCs (GFP - PDGFRα + , white arrow-head), suggesting the lack of OPC competition. Scale bar: 50 μm G. Representative images showing the expansion of OPCs with EGFR mutation ( EGFRvIII ). The expanded clone of EGFRvIII -expressing OPCs excluded WT OPCs (GFP - PDGFRα + ), suggesting OPC competition. Scale bar: 50 μm. H. Summary of the sufficiency and insufficiency in driving OPC competition of all examined gliomagenic mutations in this study.

Article Snippet: To inactivate Rb function, the HPV-16 E7 fragment was amplified from plasmid p1324 HPV-16 E7 (Addgene#8643) and cloned to replace Cas9 in MSCV vector, maintaining the in-frame fusion with T2A-EGFP for accurate reporting.

Techniques: Sequencing, Standard Deviation, Knock-Out, Generated, Transduction, Clone Assay, Mutagenesis, Expressing

Journal: Virology Journal

Article Title: Reciprocal transactivation of Merkel cell polyomavirus and high-risk human papillomavirus promoter activities and increased expression of their oncoproteins

doi: 10.1186/s12985-021-01613-0

Figure Lengend Snippet: Sequences of primers used in this study

Article Snippet: The HPV16 E6 and E7 expression plasmids were purchased from Addgene (p1322 and p1324, respectively).

Techniques: Sequencing, Binding Assay, Mutagenesis

Journal: Virology Journal

Article Title: Reciprocal transactivation of Merkel cell polyomavirus and high-risk human papillomavirus promoter activities and increased expression of their oncoproteins

doi: 10.1186/s12985-021-01613-0

Figure Lengend Snippet: HPV16 E6 and E7 stimulates the transcriptional activity of the MCPyV early and late promoter. A C33A cells were transfected with 400 ng luciferase reporter plasmid and increasing amounts (400 ng or 800 ng) of expression plasmids for HPV16 E6 and E7. B HSC-3 cells were transfected with 400 ng luciferase reporter plasmid and increasing amounts of expression plasmids for HPV16 E6 and E7. C HaCaT cells were transfected with 400 ng luciferase reporter plasmid and increasing amounts of expression plasmids for HPV16 E6 and E7. Luciferase activity was corrected for the protein concentration in each sample. Each bar represents the average of three independent parallels ± SD. The transcriptional activity of the early (respectively late) promoter in the presence of empty vector pcDNA3.1 (EV) was arbitrary set as 100%. Similar results were obtained in an independent experiment. *p < 0.05; **p < 0.01; ***p < 0.001

Article Snippet: The HPV16 E6 and E7 expression plasmids were purchased from Addgene (p1322 and p1324, respectively).

Techniques: Activity Assay, Transfection, Luciferase, Plasmid Preparation, Expressing, Protein Concentration

Journal: Virology Journal

Article Title: Reciprocal transactivation of Merkel cell polyomavirus and high-risk human papillomavirus promoter activities and increased expression of their oncoproteins

doi: 10.1186/s12985-021-01613-0

Figure Lengend Snippet: Effect of mutations in E6 and E7 on their ability to transactivate the early and late promoter of MCPyV. C33A cells were transfected with 400 ng luciferase reporter plasmid containing either the early or the late promoter of MCPyV. Co-transfection was done with 400 ng of empty expression vector (EV) or expression plasmids for wild-type LT, wild-type sT or their mutants. Luciferase activity was corrected for the protein concentration in the lysate. Each bar represents the average of three independent parallels ± SD. The transcriptional activity of the early (respectively late) promoter in the presence of empty vector pcDNA3.1 (EV) was arbitrary set as 100%. Similar results were obtained in an independent experiment. *p < 0.05; **p < 0.01; ***p < 0.001

Article Snippet: The HPV16 E6 and E7 expression plasmids were purchased from Addgene (p1322 and p1324, respectively).

Techniques: Transfection, Luciferase, Plasmid Preparation, Cotransfection, Expressing, Activity Assay, Protein Concentration

Journal: Virology Journal

Article Title: Reciprocal transactivation of Merkel cell polyomavirus and high-risk human papillomavirus promoter activities and increased expression of their oncoproteins

doi: 10.1186/s12985-021-01613-0

Figure Lengend Snippet: Effect of E6 and E7 on the transcriptional activity of MCPyV early and late promoter deletion mutants in C33A cells. A Schematic presentation of the full length and truncated MCPyV early and late promoter. The non-coding region (NCCR) consists of 464 bp was cloned in late to early direction upstream of the luciferase gene (= early MCPyV promoter) or in the early to late direction upstream of the luciferase gene (= late MCPyV promoter). The number of nucleotides in the promoter is given in parenthesis. B Cells were co-transfected with 400 ng luciferase reporter plasmid containing the MCPyV early (respectively late) promoter or truncated versions and 400 ng of expression plasmids for E6 or E7. The figure in the left panel shows transactivation by E6 or E7. The transcriptional activity of the early (respectively late) promoter in the presence of empty vector pcDNA3.1 (EV) was arbitrary set as 100%. The figure in the right panel represents the activity of early promoter (respectively late promoter) and its truncated versions. The activity of the full-length MCPyV early promoter (respectively late promoter) was arbitrary set as 100%. Luciferase activity was corrected for the protein concentration in the lysate. Each bar represent the average of three independent parallels ± SD. Similar results were obtained in an independent experiment. *p < 0.05; **p < 0.01; ***p < 0.001

Article Snippet: The HPV16 E6 and E7 expression plasmids were purchased from Addgene (p1322 and p1324, respectively).

Techniques: Activity Assay, Clone Assay, Luciferase, Transfection, Plasmid Preparation, Expressing, Protein Concentration

Journal: Virology Journal

Article Title: Reciprocal transactivation of Merkel cell polyomavirus and high-risk human papillomavirus promoter activities and increased expression of their oncoproteins

doi: 10.1186/s12985-021-01613-0

Figure Lengend Snippet: HPV16 E6 and E7 stimulate expression of MCPyV LT. MCPyV-positive WaGa cells were transfected with empty vector (EV) or plasmids coding for HPV16 E6 or HPV16 E7. Cell lysates were prepared 24 h after transfection and LT expression levels were analyzed by western blotting. GAPDH was used as a loading control. The molecular mass marker (in kDa) is shown in the lane on the left

Article Snippet: The HPV16 E6 and E7 expression plasmids were purchased from Addgene (p1322 and p1324, respectively).

Techniques: Expressing, Transfection, Plasmid Preparation, Western Blot, Control, Marker