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. Author manuscript; available in PMC: 2016 Jan 1.
Published in final edited form as: Virology. 2014 Nov 14;474:110–116. doi: 10.1016/j.virol.2014.10.013

Figure 2. Mutation in VP2/3 NLS attenuates infectivity of BKPyV.

Figure 2

(A) Equal genome numbers of wild type (Dun) and NLS mutant (DunM) viruses were separated by SDS-PAGE and analyzed by Western blotting for the presence of VP2 and VP3. (B) RPTE cells were infected with Dun and DunM virus at an MOI of 104 genomes per cell for 24 h. Whole cell lysates were resolved by SDS-PAGE and probed for TAg and β actin. Quantification was performed using the Li-Cor Odyssey system, with mock and mutant TAg levels normalized to Dun, set to a value of 1. Results represent at least three independent experiments. (C) RPTE cells were infected with equal genomes and harvested immediately or 24 h after adsorption under alkylating conditions, then whole cell lysates were resolved under reducing or non-reducing conditions by SDS-PAGE and probed for VP1 and GAPDH. (D) RPTE cells were infected with equal genomes and left untreated or treated with BFA as a control for non-specific staining, then fixed at 24 hpi and stained with anti-VP2/3 and DAPI.