Figure 5.

HuR does not seem to bind to HIV-1 RNA. A. Sequence of HIV WT ARE sequence and silent mutations introduced in the AREmut virus. B. HEK293T cells were transfected with WT or AREmut pNL4.3 proviruses. Quantification, by ELISA, of the virions produced in the cell culture supernatant, based on the detection of Gag CA-p24. C. Jurkat cells were infected with WT or AREmut NL4.3 viruses. Viral replication was monitored by ELISA quantification of HIV Gag CA-p24 antigen in the cell culture supernatant. D. Immunoprecipitation was carried out with anti-HuR antibodies or unrelated anti-HA antibodies. Western blot analysis, showing the immunoprecipitated protein. E. Coimmunoprecipitated mRNAs were detected by quantitative RT-PCR, using primers against HIV-1 gag-pol, ptma (as a positive control for HuR binding), and the housekeeping gene gapdh (as a negative control).