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Figure 3 | Retrovirology

Figure 3

From: The role of A-kinase anchoring protein 95-like protein in annealing of tRNALys3to HIV-1 RNA

Figure 3

Decreased cellular HAP95 does not affect the incorporation of viral RNA and tRNALys3into HIV-1 particles. Total viral RNA was isolated from equal amounts of purified HIV-1 particles. The abundance of viral RNA or tRNALys3 was determined by hybridizing dot blots of total viral RNA with 32P-labeled DNA probes specific for HIV-1 RNA or tRNALys3 and quantitating radioactive signals using a PhosphorImager instrument. The results were normalized to that of virions produced from siRNACon-treated cells and presented in percentage. (A) Upper panel: Diagram of HIV-1 genomic organization and the positions of 3 DNA probes used in dot blot hybridization. Numbers indicate the nucleotide positions relative to the start site (+1) of transcription of HIV-1 genomic RNA. Lower panel: A representative of dot blots probed with 3 DNA probes in triplicate (left part, 1, 2, and 3) and a graph showing the relative abundance of viral genomic RNA in HIV-1 virions (right part). m, samples obtained from mock-transfected cells. (B) A representative of dot blots probed with tRNALys3 probe in triplicate (upper panel, 1, 2, and 3) and a graph showing the relative abundance of tRNALys3 in HIV-1 virions (lower panel). (C, D) Quantitation of viral RNA and tRNALys3 in HIV-1 particles by real-time RT-PCR. Shown are the averages of fold change ± SD of 3 experiments.

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