PDE8A supports HIV-1 replication at the level of reverse transcription.

<p>(A) HEK293T cells were transfected with different concentrations of pCMV6-PDE8A or an empty vector control. PDE8A expression was analyzed forty-eight hours after transfection by western blot. (B) HEK293T cells were transfected with different concentrations of pCMV6-PDE8A or an empty vector control and infected with VSV-G/NL4-3.Luc 48 hours after transfection. Luciferase activity was measured 2 days after infection (n = 3, mean and SD). (C) HEK293T cells were transfected with different concentrations of pCMV6-PDE8A or an empty vector control and infected with VSV-G/NL4-3-Ba-L 48 hours after transfection. Proviral DNA levels were analyzed forty-eight hours after infection by qPCR. Proviral DNA levels are expressed relative to the control after correction for β-actin levels in the same sample (n = 3, mean and SD). (D) HEK293T cells were transfected with 1 µg of pCMV6-PDE8A or 1 µg of empty vector control and infected with VSV-G/NL4-3-Ba-L 48 hours after transfection. <i>R/U5</i> and <i>Pol</i> proviral DNA levels were analyzed by qPCR during 48 hours post-infection. <i>R/U5</i> and <i>Pol</i> levels are expressed as the ratio of the expression in cells transfected with pCMV-PDE8A divided by the expression in cells that were transfected with the empty vector control and corrected for β-actin levels (n = 2, mean and SD). (E) HEK293T cells were transfected with 1 µg of pCMV6-PDE8A or 1 µg of the empty vector control and infected with VSV-G/NL4-3-Ba-L 48 hours after transfection. Integrated HIV-1 copies were determined 48 hours post infection by Alu-PCR and expressed relative to the control after correction for β-actin levels in the same sample (n = 2, mean and SD). Statistical differences were analyzed using an un-paired T test: *p<0.05, **p<0.01, ***p<0.001.