Fig. 1

Dcr2 interacts with the N-terminal region of Symplekin a Dcr2 co-immunoprecipitates (co-IP) with the core cleavage complex (CCC) and R2D2 in Drosophila culture cell crude nuclear extract. Antibodies used for immunoprecipitation (IP) are shown above. Antibodies used for western blot (WB) are listed to the left. ‘Beads’ and ‘α-Myc’ are negative controls. No sample was loaded in lanes labeled ‘None.’ b Dcr2 binds Symplekin amino acids 1-271 and not amino acids 272-1165. Exogenously expressed HA-tagged Symplekin deletions are defined above the blots. Other labels are as in a. WB of full length Symplekin and Symplekin mutant IPs are the top figure while co-IP of Symp(1-271) with Dcr2 is shown in the bottom WB. c Dcr2 binds exogenously expressed CCC components CPSF73 and CPSF100. HA-tagged, full-length CPSF73 and CPSF100 were IPed from Drosophila culture cells stably expressing these proteins. Dcr2 co-IPs with both CPSF73 and CPSF100 was identified by western blot (top). Other labels are as in a. Dcr2 was IPed from these cells. Co-IP of HA-CPSF73 and HA-CPSF100 was identified by western blot (bottom). Controls are as in a. d CCC components CPSF100 and CPSF73 do not interact with Dcr2 in the absence of full length Symplekin. WB of Symp(1-271) IP (top) and WB of Symp (272-1165) IP (bottom) from Symplekin RNAi-depleted samples are shown. WB are labeled as in a