Fig. 3
Transduction of Daudi B-cells by HA-Vpr-delivering VLPs increases genome uracilation. Daudi B-cells were transduced with VLP HA-Vpr at a MOI of 5. a UNG2 levels were analyzed by western blot with anti-UNG2 antibodies at 24 and 120 h post infection. Lanes were cropped from blot membranes serially probed with anti-UNG2 then with anti-αtubulin antibodies (see Additional file 2: Supplementary dataset 2). UNG activity was measured in the corresponding whole cell extracts. b DNA was extracted from cells 120 h post-transduction and treated with Methoxyamine (Mx) to protect pre-existing AP sites. Recombinant E. coli UDG was then used to excise uracil and generate AP sites which were quantified by ELISA. Numbers of AP sites per 105 bp were measured in triplicate with the error bars representing the standard error of the mean. c DNA of ethanol-fixed cells was labeled with propidium iodide. Cell cycle progression was followed by flow cytometry. G2/M:G0/G1 ratios are indicated in the upper right corner of each histogram. d Apoptosis was followed by flow cytometry after labeling with Annexin V-FITC in the presence of CaCl2. Ut: untransduced