Fig. 1

HA-Vpr-delivering VLPs decrease UNG2 expression in human B lymphocytes. a Daudi cells were infected with VSV-G pseudotyped wt HIV-1 at MOI of 25. 24 and 72 h after infection UNG2 levels were examined by immunoblot with anti-UNG2 antibody or anti-α-tubulin as loading control. b Daudi-CD4 + cells were infected with either HIV-1 wt, HIV-1ΔVpr or VSV-G pseudotyped wt HIV-1 at the indicated MOI. 72 h after infection UNG2 levels were examined by immunoblot with anti-UNG2 antibody or anti-α-tubulin as loading control (Ni: non-infected). c HA-Vpr-delivering VLPs were generated by cotransfecting HEK 293T cells with pCMV HA-Vpr along with a packaging vector pCMVΔR8.2ΔVpr, an Envelope vector pCMV VSV-G and the defective lentiviral vector pHR-GFP. VLPs produced in the presence (VLP HA-Vpr) or absence of pCMV HA-Vpr (VLP ΔVpr) were purified on sucrose gradients, and titrated by p24 ELISA kit. VLP titers were also determined by transducing 293T cells with serial dilutions of virus suspensions. Virus extracts corresponding to the indicated p24 amount were resolved on PAGE-SDS gels, and immunoblotted with specific antibodies for their content in p24 and HA-Vpr. d Daudi B-cells were transduced with VLP HA-Vpr or ΔVpr at M.O.I of 10. Cells were lysed at different time points and examined by immunoblot for UNG2 content with anti-UNG2 antibody or anti-α-tubulin as loading control. Lanes were regrouped from two different gels, see Supplementary dataset for detail. UNG2 levels at the various time points were quantified with ImageJ and normalized with their corresponding α-tubulin levels and plotted as a UNG2% of untransduced (Ut) cells. Lanes were cropped from blot membranes serially probed with anti-UNG2 then with anti-αtubulin antibodies (see Additional file 2: Supplementary dataset 2)