Journal: Molecular Oncology

Article Title: Preclinical evaluation of the gorilla‐derived HAdV‐B AdV ‐lumc007 oncolytic adenovirus ‘ GoraVir ’ for the treatment of pancreatic ductal adenocarcinoma

doi: 10.1002/1878-0261.13561

Figure Lengend Snippet: Infection of MIA PaCa‐2 CD46 knockout cells with GoraVir and HAdV‐C5. (A) Cell surface protein expression of CAR, CD46, and DSG‐2 on unstained (−), wildtype (WT), empty vector (EV) control, or CD46 knock out (KO) MIA PaCa‐2 cells; (B) WT, EV, and KO MIA PaCa‐2 cells were infected with GoraVir or HAdV‐C5 at multiplicity of infection (MOI) 10 for 48 h after which hexon protein expression was measured by flow cytometry. Depicted are representative figures of n = 3 biological replicates; (C) Percentage of hexon positive cells relative to WT‐infected cells. Depicted are means of n = 3 biological replicates; (D) WT, EV, and KO MIA PaCa‐2 cells were infected with GoraVir or (E) HAdV‐C5 at different MOI and cell viability was measured by WST assay at 6 days post infection. Depicted are mean ± SEM of n = 3 independent experiments each performed in triplicate.

Article Snippet: Cells were stained extracellular with primary antibodies anti‐CAR (1 : 1000, #05‐644, Millipore, Amsterdam, The Netherlands), anti‐CD46 (1 : 125, #555948, BD), or DSG‐2 PE (1 : 150, #12‐9159‐42, Invitrogen) according to standardized methods.

Techniques: Infection, Knock-Out, Expressing, Plasmid Preparation, Flow Cytometry, WST Assay