Figure 1

EU staining of HIV-1 RNA in vitro . (A) EU-labeled virus encoding MS2-binding sites and produced in the presence of MS2-GFP (green) was treated with cell extraction buffer, 293T cell lysate, or 293T cell lysate containing rhTRIM5α for 15 minutes, fixed onto slides, and then stained for EU (red). Particles were visualized by fluorescent confocal microscopy and shown for each individual color and with both colors (merge). (B) MS2-GFP+ and EU+ virions were counted per field after treatment with buffer, 293T cell extract, or 293T cell extract expressing rhTRIM5α in the presence of 10 μg/ml RNase A. Data represent the ratio of double positive particles of the total counted MS2-GFP+ particles ± SEM of at least 4 fields. Results are representative of 2 independent experiments. A student’s t test was performed on medium vs. RNase A treatment of virions incubated with either 293T or 293T-rhTRIM5α lysate. NS denotes a non-significant p value (>0.05).