Fig 2.
(A) ORFV073 protein expression kinetics. OFTu cells were mock infected or infected with ORFV073 revertant virus (OV-IA82RV073Flag) (MOI = 10) for 2, 4, 6, 8, 10, 12 and 24 h p.i. Whole cell protein extracts (50 μg) were resolved by SDS-PAGE, blotted to nitrocellulose membranes and probed with antibodies against flag and GAPDH (control). (B) Subcellular localization of ORFV073 (top panels) and structural protein ORFV086 (bottom panels) in OFTu cells mock infected or infected with revertant virus OV-IA82RV073Flag or wild type virus OV-IA82, respectively (MOI = 10). Cells were fixed at 30 min and 1, 2, 6, 8, 12, 16 and 24 h p.i., incubated with anti-flag or anti-ORFV086 monoclonal antibodies, sequentially stained with Alexa Fluor 488-labeled secondary antibody and DAPI, and examined by confocal microscopy. Insets show magnified areas of the field. Shown are representative images from 24 h p.i. ORFV073 and ORFV086, green; DAPI, blue. Results are representative of two independent experiments.