Fig 3.

The replication defect observed in N121K virus occurs at the nuclear import stage. HeLa cells (2 × 10⁵) were infected with 4 ng of p24 from vesicular stomatitis virus G protein-pseudotyped WT, A92E, or N121K virus (harboring the luciferase gene). The infection was performed under the following conditions: no drug, 1 μM CsA, 1 μM azidothymidine (AZT; Sigma-Aldrich), and 1 μM raltegravir (RAL). (A) Virus infectivity was determined by measuring the luciferase activity 2 days after infection. Total DNA was extracted 24 h (B and C) or 36 h (D) after infection, and 200 ng of the DNA was subjected to quantitative PCR (19). The error bars represent the standard deviations from duplicate assays performed in a single experiment. The results are representative of two independent experiments. UD, undetectable.