Figure 1

Identification of proteins that co-immunoprecipitate with Zn72D. (A) IPs were performed with an anti-HA antibody in wild-type (wt) S2 cells and in S2 cells expressing HA-Zn72D. Proteins that co-IP with HA-Zn72D were identified by mass spectrometry. (B) (Top) HA-Zn72D co-IPs with Myc-CG5641 when cells expressing HA-Zn72D were transiently transfected with a vector expressing Myc-CG5641 (left 2 lanes). IPs were performed using an anti-Myc antibody, and immunoblotting was performed with an HA antibody. Right 2 lanes are controls in which no Myc-CG5641 was transfected. (Bottom left) HA-Zn72D co-IPs with FMR1. IPs were performed using an anti-FMR1 antibody and immunoblotting was performed to detect HA. IPs were performed in cells that did (2 left lanes) and did not (2 right lanes) express HA-Zn72D. (Bottom right) The FMR1 antibody specifically recognizes FMR1 protein. S2 cell extracts from untreated and FMR1 knockdown cells were probed with anti-FMR1 and anti-γ-tubulin. (C) The interaction between Zn72D and Bel is RNA-independent. IPs using anti-HA antibody were performed on S2 cells expressing HA-Zn72D, in the absence (lane 3) or presence (lane 6) of RNase A. Lane 9 is a control IP performed with an anti-HA antibody using S2 cells not expressing HA-Zn72D. HA and Bel were detected using immunoblotting. The ethidium bromide stained gel to the right shows equivalent amounts of input extract loaded per lane, either untreated or treated with RNase A. For parts B and C, "in" indicates 2% of the input into each IP and "FT" indicates 2% of flow-through from each IP.