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Figure 4 | Retrovirology

Figure 4

From: Intracytoplasmic maturation of the human immunodeficiency virus type 1 reverse transcription complexes determines their capacity to integrate into chromatin

Figure 4

Analysis of protein composition of cytoplasmic and nuclear RTCs. cRTCs and nRTCs purified 5 h after infection were immunoprecipitated using the indicated antibodies and Protein G Sepharose. DNA was isolated from immune complexes and analyzed by real-time PCR as in Fig. 1. DNA recovered in immunoprecipitated RTCs as percentage of total HIV-1 DNA detected in the cRTCs is indicated under the histogram columns. DNA recovery for isotype control antibodies is shown on the right. DNA recovery for mouse mAb is shown in open boxes, for rabbit polyclonal antibodies – in shaded boxes. A,B. Immunoprecipitated cRTCs were analyzed using primers specific for early (A) and late (B) reverse transcription products. N.d. – not done. Results are mean ± SD of triplicate determinations, except for late DNA analysis of anti-MA-precipitated complexes, which was done only once. One representative experiment out of 4 performed is shown. C. Experiment was performed as in A, except that nRTCs were analyzed. Low sensitivity of primers specific for late HIV-1 DNA precluded their use for analysis of nRTCs. Results are mean ± SD of triplicate determinations. One representative experiment out of 4 performed is shown. D. Temporal analysis of cRTCs. Results are mean ± SD of triplicate determinations. One representative experiment out of 3 performed is shown.

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Retrovirology

ISSN: 1742-4690

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