Journal: Scientific Reports

Article Title: The oncolytic adenovirus VCN-01 promotes anti-tumor effect in primitive neuroectodermal tumor models

doi: 10.1038/s41598-019-51014-1

Figure Lengend Snippet: Characterization of VCN-01 in PNET cell lines in vitro . ( A ) Expression of adenoviral receptors CAR, α v β 3 integrin and α v β 5 integrin in CNS-PNET cell lines PFSK-1 (left) and SK-PN-DW (right). Graph shows the percentage of stained cells for each receptor (Mean ± SD; n = 3). ( B ) PFSK-1 and SK-PN-DW (200,000 cells) infected with the GFP expressing vector AdTLRGDK at MOIs of 0, 0.1, 1, 10 or 100 PFU/cell. Graph indicates the percentage of GFP positive cells expression measured by flow cytometry at 24 h (white bars) or 48 h (black bars) after the infection (Mean ± SD; n = 3). ( C ) Detection of the viral proteins E1A and fiber in whole-cell lysates 48 h after being from PFSK-1 and SK-PN-DW from VCN-01 infected PFSK-1 and SK-PN-DW cultures. Grb2 was used as loading control protein. Blots from different parts of the same gel have been grouped to improve clarity of the image. ( D ) Viral titers in PFSK-1 and SK-PN-DW (50,000 cells/well) cultures 72 h after being infected with VCN-01 at MOIs 1 and 10. Bars represent total PFUs contained in the lysates (Mean ± SD; n = 3).

Article Snippet: PNET cell lines PFSK-1 (ATCC, Manassas, VA, USA; CRL-2060 TM ) and SK-PN-DW (ATCC, CRL-2139 TM ) were stained with unlabeled monoclonal antibodies recognizing the adenoviral receptors CAR (Merck Millipore, Temecula, CA, USA; 05–644) α v β 3 integrin (Merck Millipore; CBL544) and α v β 5 integrin (R&D Systems Inc., Minneapolis, MN, USA; MAB2528), and donkey anti-mouse IgG1-AlexaFluor488 (Thermo Fisher Scientific, Waltham, MA, USA; A-21202) as secondary antibody.

Techniques: In Vitro, Expressing, Staining, Infection, Plasmid Preparation, Flow Cytometry, Control