FIG. 2.

Southern blot of scAAV vector DNA from normal and ATM^−/− cells. (a) Diagrams of intact GFP and GFP-CD vector genomes with relevant restriction sites indicated. (b) Southern blot of uncut low-molecular-weight DNA extracted from cytoplasmic (C) and nuclear (N) fractions from vector-infected or control uninfected wt and ATM^−/− cells. Cells were infected at an MOI of 250 with intact GFP (scAAV-GFP) or circularization-dependent GFP (scAAV-GFP-CD) vectors and harvested at 24 h postinfection. The position of the linear vector genome is indicated (2,432 bp). (c) Southern blot of nuclear fractions from the same infections as described above. Nuclear fractions from wt or ATM^−/− cells were divided into four aliquots and digested with XbaI (X), MluI (M), or Plasmid Safe nuclease (P) or were left undigested (U). The sizes in bp of the intact linear (2,432 bp) and digested vector DNA bands are indicated.