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Figure 3 | Retrovirology

Figure 3

From: HuR interacts with human immunodeficiency virus type 1 reverse transcriptase, and modulates reverse transcription in infected cells

Figure 3

HuR is involved in the early steps of HIV-1 replication cycle. A. siRNA silencing of HuR revealed by Western blot. HeLa P4.2 cells (CD4+, LTR-LacZ) were transfected with 30 nM of siRNAs directed against HuR (HuR1, HuR2, HuR3), H2O, or a non targeting siRNA (Ctrl). For each siRNA, five wells were infected with the HIV-1NL4.3 strain, 72 hours after transfection with the siRNA. The contents of one well were collected and lysed, to check that HuR expression was silenced at the time of infection (A, upper panel). B. Effect of HuR silencing on the infection of cells by wild type HIV-1. 24 hours post-infection, infected cells were counted after the fixation and X-Gal staining of triplicate wells. The contents of one well were collected and lysed, to check that HuR expression was effectively silenced at the time of fixation (A, lower panel). The results presented are a compilation of six independent experiments, normalized as a function of the results obtained with the control siRNA (Ctrl). C. Effect of HuR silencing on HIV-1 reverse transcription. HeLa cells were treated with siRNA HuR-1 or a non targeting siRNA (Ctrl), then infected with HIV-1ΔEnv-luciferase VSV-G pseudotyped viruses at an MOI of 1. Total DNA was extracted from the infected cells and RT products were quantified by quantitative real-time PCR, 16 hours after infection. ssRT: minus-strand strong stop DNA, flRT: full-length HIV DNA, HBB, human beta-globin. D. Effect of HuR overexpression on HIV-1 reverse transcription. As in C, except that the cells were transfected with a vector allowing HuR overexpression (pCMV-HuR) or an empty vector (pcDNA3), before infection.

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