Journal: Cellular Microbiology

Article Title: Changes in nucleolar morphology and proteins during infection with the coronavirus infectious bronchitis virus

doi: 10.1111/j.1462-5822.2006.00698.x

Figure Lengend Snippet: A. Distribution of p53 in mock‐infected cells. B. Confocal images of cells expressing IBV proteins (green) and showing the distribution of p53 (red) and merged at 24 h pi. C. Enlarged images of IBV‐infected cells with IBV proteins labelled in green and p53 in red at 24 h pi. Transmission phase contrast (TPC) of the lower cell is shown below. Nucleoli are indicated (No) and the size bar is 10 µm. D. Western blot analysis of p53 in mock and infected cells at 0 and 24 h pi. Protein concentrations were standardized and compared with GAPDH.

Article Snippet: To detect the tumour suppressor protein p53 we used a mouse anti‐p53 antibody (D01, Santa Cruz) and to detect GAPDH we used a mouse anti‐GAPDH antibody (6C5, AbCam), with appropriate secondary antibody.

Techniques: Infection, Expressing, Transmission Assay, Western Blot

Journal: Cellular Microbiology

Article Title: Changes in nucleolar morphology and proteins during infection with the coronavirus infectious bronchitis virus

doi: 10.1111/j.1462-5822.2006.00698.x

Figure Lengend Snippet: A. Confocal analysis to investigate the potential colocalization of p53 (red) in cells expressing EGFP‐IBV N protein (green). B and C. Confocal analysis of the distribution of IBV proteins (blue), EGFP‐IBV N protein (green) and p53 (red) in IBV‐infected cells at 24 h pi. Colocalization between EGFP‐IBV N protein plus p53, EGFP‐IBV N protein plus IBV proteins, p53 plus IBV proteins, and between the three complexes, appears yellow, cyan, purple and white respectively. The nucleolus is indicated (No). Size bar is 10 µm.

Article Snippet: To detect the tumour suppressor protein p53 we used a mouse anti‐p53 antibody (D01, Santa Cruz) and to detect GAPDH we used a mouse anti‐GAPDH antibody (6C5, AbCam), with appropriate secondary antibody.

Techniques: Expressing, Infection

Journal: bioRxiv

Article Title: BK polyomavirus (BKPyV) is a risk factor for bladder cancer through induction of APOBEC3-mediated genomic damage

doi: 10.1101/2021.05.13.443803

Figure Lengend Snippet: Schematic model of BKPyV hit-and-run carcinogenesis hypothesis. Immune-insufficiency leads to sloughing of actively infected renal “decoy” cells and BKPyV viruria. BKPyV infects the G0-arrested urothelium. BKPyV LT-Ag inhibits retinoblastoma (pRb) and p53 inhibition of the cell cycle. BKPyV cell cycle re-entry progresses to arrest at the G2/M checkpoint. BKPyV stimulates APOBEC3 enzyme activity and host genome damage that inactivates tumour suppressors. The immune system clears the virus but initiated cells persist and over a period of years expand to form a tumour.

Article Snippet: Primary antibodies, applied overnight at 4°C, were anti-SV40 LT-Ag (1:200, mouse monoclonal “Pab 108”, Santa Cruz Biotechnology, sc-148), anti-VP1 (1:100, mouse “pab597”, kind gift from Chris Buck at the National Institute of Health, Bethesda), anti-Rad51 (1:1,000, rabbit “EPR4030(3)”, Abcam #ab133534), anti-Ki67 (1:400, mouse, “MM1”, Leica, NCL-L-Ki67-MM1), anti-MCM2 (1:500, rabbit, “D7G11”, Cell Signalling #3619), anti-Phospho-pRb Serine 608/807/811 (1:500, rabbit, Cell Signalling #9308), anti-p53 (1:40, mouse, “D01”, Cell Signalling #18032), anti-ZO3 (1:800, rabbit, Cell Signalling #3704) and anti-APOBEC3A/B/G (1:100 rabbit monoclonal, clone 5210-87-13 ).

Techniques: Infection, Inhibition, Activity Assay, Virus

Journal: bioRxiv

Article Title: BK polyomavirus (BKPyV) is a risk factor for bladder cancer through induction of APOBEC3-mediated genomic damage

doi: 10.1101/2021.05.13.443803

Figure Lengend Snippet: Homologous recombination was induced by BKPyV infection of human urothelium. (A) Western blotting for p53 protein showed significant stabilisation (no increase in transcription was observed) during BKPyV infection. (B) However, there was no increased expression of verified p53 target genes . RAD51 and (D) RAD51AP1 transcripts were significantly induced by BKPyV infection. (E) Western blotting confirmed significant Rad51 protein induction and phosphorylation was observed as a slight increase in molecular weight when Rad51 was induced by BKPyV. (F) Analysis of indirect immunofluorescence for Rad51 found nuclear speckles were significantly increased in BKPyV infection. In some cells, indirect immunofluorescence revealed LT-Ag and Rad51 protein colocalisation to large granular deposits in the nuclei of BKPyV-infected urothelial cells. Scale bar denotes 10 μm, immunofluorescence performed on n=3 independent donors with representative images shown. Full blots and further immunofluorescence images for p53 and Rad 51 in Extended Data Fig. 17 & 18, respectively.

Article Snippet: Primary antibodies, applied overnight at 4°C, were anti-SV40 LT-Ag (1:200, mouse monoclonal “Pab 108”, Santa Cruz Biotechnology, sc-148), anti-VP1 (1:100, mouse “pab597”, kind gift from Chris Buck at the National Institute of Health, Bethesda), anti-Rad51 (1:1,000, rabbit “EPR4030(3)”, Abcam #ab133534), anti-Ki67 (1:400, mouse, “MM1”, Leica, NCL-L-Ki67-MM1), anti-MCM2 (1:500, rabbit, “D7G11”, Cell Signalling #3619), anti-Phospho-pRb Serine 608/807/811 (1:500, rabbit, Cell Signalling #9308), anti-p53 (1:40, mouse, “D01”, Cell Signalling #18032), anti-ZO3 (1:800, rabbit, Cell Signalling #3704) and anti-APOBEC3A/B/G (1:100 rabbit monoclonal, clone 5210-87-13 ).

Techniques: Homologous Recombination, Infection, Western Blot, Expressing, Phospho-proteomics, Molecular Weight, Immunofluorescence

Journal: bioRxiv

Article Title: BK polyomavirus (BKPyV) is a risk factor for bladder cancer through induction of APOBEC3-mediated genomic damage

doi: 10.1101/2021.05.13.443803

Figure Lengend Snippet: Schematic of hypothetical model of urothelial DNA damage at displacement loops at the G2 checkpoint during BKPyV infection. LT-Ag interactions might derive from (A) its DNA-binding helicase activity or (B) one of the other dimerization domains present in the LT-Ag protein. (C) LT-Ag inhibition of p53 is likely important for this process as p53 would normally prevent Rad51 oligomerisation by direct binding . The evidence from this study is most robust for BKPyV-induced APOBEC3B activity but APOBEC3A protein was consistently more abundant during infection and its role requires further validation.

Article Snippet: Primary antibodies, applied overnight at 4°C, were anti-SV40 LT-Ag (1:200, mouse monoclonal “Pab 108”, Santa Cruz Biotechnology, sc-148), anti-VP1 (1:100, mouse “pab597”, kind gift from Chris Buck at the National Institute of Health, Bethesda), anti-Rad51 (1:1,000, rabbit “EPR4030(3)”, Abcam #ab133534), anti-Ki67 (1:400, mouse, “MM1”, Leica, NCL-L-Ki67-MM1), anti-MCM2 (1:500, rabbit, “D7G11”, Cell Signalling #3619), anti-Phospho-pRb Serine 608/807/811 (1:500, rabbit, Cell Signalling #9308), anti-p53 (1:40, mouse, “D01”, Cell Signalling #18032), anti-ZO3 (1:800, rabbit, Cell Signalling #3704) and anti-APOBEC3A/B/G (1:100 rabbit monoclonal, clone 5210-87-13 ).

Techniques: Infection, Binding Assay, Activity Assay, Inhibition, Biomarker Discovery

Journal: BioMed Research International

Article Title: Alternative Splicing Generates Different Parkin Protein Isoforms: Evidences in Human, Rat, and Mouse Brain

doi: 10.1155/2014/690796

Figure Lengend Snippet: List of antibodies targeting PARK2 isoforms.

Article Snippet: H00005071-D01 , Abnova.

Techniques: