Figure 3.

MARF1 NYN domain is required to degrade a target mRNA. (A) Schematic diagram of the basic Renilla luciferase-encoding mRNA reporter, containing five 19-nt BoxB hairpins, interacting with λN-HA-MARF1. (B) Schematic diagram of full-length MARF1 and MARF1 fragments used in tethering assays. (C) Expression levels of λNHA-tagged fusion proteins, as determined by western analysis using anti-HA and anti-actin antibodies. (D) RL activity detected in extracts from HeLa cells expressing the indicated proteins. Cells were cotransfected with constructs expressing the RL-5BoxB reporter, FL, and indicated fusion proteins. Histograms represented normalized mean values of RL activity and mRNA levels from a minimum of three experiments. RL activity values seen in the presence of λNHA-LacZ were set as 100. (E) Representative Northern blot of RL-5BoxB and FL mRNAs levels.