FIG 4.

GFZF is a transcriptional coactivator. (A) Overview of the luciferase assay used to test GFZF's role in transcription activation. S2R+ cells were treated with dsRNA targeting either lacZ (RNAi control), exon 2 of GFZF, or the 5′ UTR of endogenous GFZF. The exon 2 RNAi targets both the endogenous and transfected versions of GFZF, and the 5′ UTR RNAi targets only the endogenous GFZF. After 24 h of incubation with dsRNA, luciferase reporter plasmids and a vector that expresses either empty or wild-type GFZF were transfected into the cells. Cells were incubated for an additional 48 h and then assayed in tandem for firefly and Renilla luciferase activity. Firefly luciferase expression was driven by a GFZF-bound promoter, while Renilla luciferase expression was driven by the GFZF- and M1BP-less RpIII128 promoter. (B to E) Firefly/Renilla luciferase ratios are displayed for RpLP1, Sxl, roX2, and awd. All values are normalized to the lacZ dsRNA, empty-vector sample at each promoter.