Figure 2
Enhanced early HIV-1 NL4-3 and HIV-2 ROD-A gene expression in primary T-cells from hCycT1-tg rats after nucleofection of proviral DNA. Activated T-cells from hCycT1-tg and n-tg rats were nucleofected with proviral GFP reporter constructs pHIV-1NL4-3 GFP or pHIV-2ROD-A GFP in principle as reported [3, 27]. The GFP expression level in viable cells was analyzed 24 h later by flow cytometry. (A) Representative flow cytometry dot plots of nucleofected T-cells. Living cells were identified by their forward scatter (FSC) and side scatter (SSC) characteristics (left panels, R1 gate). The GFP fluorescence in living cells was analyzed against an empty reference channel (right panels, FL-4). The MFI of GFP-expressing cells (right panels, R2 gate) was determined as a surrogate marker for early viral gene expression. (B) Cumulative results from several independent experiments. Each closed circle depicts the MFI of provirus-nucleofected T-cells as the mean of triplicates performed for cultures from individual animals. Open triangles represent the arithmetic mean of the MFI of all animals in one group ± SEM. The indicated p-values were calculated using the unpaired Student's t-test.