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Figure 1 | Retrovirology

Figure 1

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

Figure 1

Analysis of nucleo-cytoplasmic distribution of HIV-1 RTCs.HeLa cells were spinoculated with MLV Env-pseudotyped NL4-3 or NL4-3-GFP HIV-1. A. HeLa cells infected with GFP-expressing HIV-1 were analyzed by FACS 48 h after infection. Percentage of GFP-positive cells was counted using CellQuest software. B. PCR analysis of the purity of nuclear extracts. Cytoplasmic and nuclear extracts were prepared from the same number of cells (1 × 106) and total DNA was isolated. Undiluted and diluted (1:10, 1:102, 1:103, and 1:104) DNA samples were analyzed by PCR using primers specific for mitochondrial DNA. M – DNA molecular size marker, NC – negative control (H2O). C,D. Real-time PCR analysis of nuclear and cytoplasmic RTCs. DNA isolated from cytoplasmic and nuclear RTCs 2 h (C) and 5 h (D) after spinoculation was analyzed in triplicate with primers specific for early or late HIV-1 DNA using SYBR Green qPCR. Serial dilutions of DNA from 8E5 cells were used as quantitative standards. Results are presented as mean ± SD.

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