Skip to main content
. 2010 Jan;16(1):170–185. doi: 10.1261/rna.1873910

FIGURE 7.

FIGURE 7.

Confirmation of the production of the γHV68 miRNAs by RNA pol III using the Left End plasmids. (A) Schematic of the γHV68 RNA pol III promoter deletion strategy. (B) RLM-RT-PCR for miRNAs in 293 cells transfected with either pLE-WT or the pLE-KO plasmids. hsa-miR-15a serves as an endogenous control for miRNA expression in 293 cells. RT-PCR products were resolved by gel electrophoresis and visualized by ethidium bromide. (C) Results from the dual luciferase reporter analysis following transfection with either the pLE-WT or pLE-KO plasmid. pGL3 luciferase reporter target is labeled below the graph. All results are displayed as a ratio of firefly luciferase expression in the pLE-WT transfected cells divided by firefly luciferase expression in the pLE-KO transfected cells. Firefly luciferase readings are all normalized to the renilla luciferase transfection control. Readings obtained from 3T3 transfected cells are shown in solid black bars; 293 cells, in solid white bars. Readings below the dashed line represent repression by the corresponding viral miRNA in pLE-WT transfected cells.