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. 2018 Apr 13;92(9):e01844-17. doi: 10.1128/JVI.01844-17

FIG 1.

FIG 1

uc002yug.2 enhances HIV-1 replication. (A to D) uc002yug.2 overexpression enhances HIV-1 replication in HeLa cells. (A) The pNL4-3 viral vector with increasing amounts of uc002yug.2 (0, 100, 300, and 900 ng) was cotransfected into HeLa cells as indicated. After 48 h, cells and supernatants were harvested. The RNA level of uc002yug.2 was detected with the qRT-PCR assay. (B) p55 levels in cell lysate and CAp24 levels in supernatants were determined by immunoblotting, and the densities of bands were analyzed with ImageJ software to calculate the values relative to that for β-actin. (C) The infectivity of HIV-1 was measured by detecting the luciferase activity after TZM-bl cells were infected with the supernatant for another 48 h. (D) The mRNA levels of RUNX1a and RUNX1b and -1c were detected by qRT-PCR in the same cells as for panel A. (E to H) Knockdown of uc002yug.2 decreases HIV-1 replication in HEK293T cells. The endogenous uc002yug.2 in HEK293T cells was knocked down by using uc002yug.2-specific shRNA. (E) The knockdown of uc002yug.2 and corresponding RUNX1a and RUNX1b and -1c mRNA levels were detected by qRT-PCR. (F) Both control and uc002yug.2sh HEK293T cells were cotransfected with pNL4-3 viral vector plus uc002yug.2 or control vector as indicated. At 48 h after transfection, the cells and supernatants were harvested. p55 levels in cell lysate and CAp24 levels in supernatants were determined by immunoblotting, and the densities of bands were analyzed with ImageJ software to calculate the values relative to that for β-actin. (G) Virus production was measured by detecting the luciferase activity after TZM-bl cells were infected with the supernatant for another 48 h. The infectivity of 293T-pLKO.1 cells transected with pNL4-3 plus control vector was set as 100%. (H) The RNA level of uc002yug.2 in the same cells as described for panel F was detected by qRT-PCR. Results are representative of those from three independent repeats, and the corresponding value of the mock control was set as 1 or 100%. Data are presented as means ± SDs.