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. 2010 May 12;5(5):e10606. doi: 10.1371/journal.pone.0010606

Figure 6. Mutation of tAg residues proline 99 and cysteine 157 reduce JCV DNA replication.

Figure 6

PHFG cells in 60 mm plates were transfected in duplicate with 400 ng of wild type (T+/t+/T'+) or 8 different mutant (muT+/t−/T'+, T+/P99At+/T'+, T+/C157At+/T'+, T+/t+/T'−, T+/t−/T'−, muT+/t+/T'+, T−/t+/T'−, T+/t−/T'+) JCV genomes. Duplicate, independent samples representing each DNA construct were extracted by the method of Hirt [39] on days 0, 7, 10 and 14 p.t. and analyzed using the Dpn1 assay as described in the legend to Figure 5. The marker (M) shown in the first lane of each blot is 1 ng of linear JCV DNA (5130 bp), and the position of Dpn1-resistent replicating genomes is denoted by an arrow at days 7, 10 and 14 p.t.