Levels of HBV DNA present in ZFN-treated HepAD38 cells and levels of secreted infectious HBV.

<p>(<b><i>a</i></b>) For experiments shown in panels <b><i>b</i></b> and <b><i>c</i></b>, HepAD38 cells in the presence of dox were transduced with ZFN- or control-expressing scAAV2 vectors (total MOI 10000 genomes/cell) and 3 days later dox was removed from culture medium to enable HBV replication. Cells were left in culture for a further 7 days before HBV genomic levels were quantified in cells (<b><i>b</i></b>) and infectious HBV levels were quantified in supernatants (<b><i>c</i></b>) by ddPCR. (<b><i>d–e</i></b>) HepAD38 cells were passaged every two to three days over the course of 14 days following treatment with ZFN- or control-expressing scAAV vectors and monitored for cellular (<b><i>d</i></b>) and supernatant (<b><i>e</i></b>) HBV levels by ddPCR. ddPCR – droplet digital polymerase chain reaction; dox – doxycycline; eGFP – enhanced green fluorescent protein; scAAV – self-complementary adeno-associated virus; UC – untreated control; ZFN – zinc finger nuclease. *p≤0.05.