Fig. 3

 GST-Stau1 fusion protein binds to oskar and nanos1 RNA. (A) SDS-PAGE analysis of purified GST fusion proteins. L, Rainbow ladder, GST Stau1 RBD4, GST, GST Stau1 RBD4 digested with thrombin protease to release GST (28 kDa) and Stau1 RBD4 (7 kDa), GST Stau2 RBD5. (B) RT-PCR of globin, gurken, or oskar cDNAs from GST Stau1 RBD4 beads. No template control, positive globin, gurken, or oskar plasmid controls, RT−, RT+, GST (1 μg) incubated with 250 ng of globin, gurken, or oskar RNA, Control Glutathione beads incubated with 250 ng of globin, gurken, or oskar RNA, and GST Stau1(1 μg) beads incubated with 250 ng of globin, gurken, or oskar RNA. (C) RT-PCR from GST Stau1 RBD4 beads incubated with zebrafish ovary RNA. No template control, positive cyclinB, actin, or nanos plasmid controls, RT−, RT+, GST protein (1 μg) incubated with 5 μg of zebrafish ovary RNA, Glutathione beads incubated with 5 μg of zebrafish ovary RNA, GST Stau1 beads (1 μg) incubated with 2.5 μg of zebrafish ovary RNA, and GST Stau1 beads (1 μg) incubated with 5 μg of zebrafish ovary RNA.