FIG. 3.

Overexpression of PRMT6 increases the steady-state level of HIV-1 provirus-expressed Tat. (A) Schematic diagram of the HIV-1 provirus in the pGCH-FLAG-Tat expression vector. The promoter is a fusion between the CMV immediate-early promoter and the R and U5 regions of the HIV-1 long terminal repeat (LTR). The CMV promoter enables high-level Tat-independent proviral-gene expression. The FLAG tag sequence (gray box) was cloned onto the 5′ end of the tat open reading frame. +1, start of proviral transcription. (B) Western blot of HeLa cells expressing pGCH-FLAG-Tat alone (−) or coexpressing Myc-PRMT6 (+). Samples were cotransfected with β-galactosidase vector, and the lysates were normalized for β-galactosidase activity. HIV antiserum was used to detect provirus expression with the 55-kDa Gag band shown.