Figure 6. NOT10 directly interacts with NOT11. (A) NOT10 and NOT11 (also known as CG13567 or C2orf29) are predicted to be primarily α-helical. NOT11 contains a conserved domain of unknown function named DUF2363. (B and C) S2 cells were transiently transfected with expression vectors encoding GFP-tagged NOT10, NOT11 or CAF40 and HA-tagged NOT10 (B) or NOT11 (C). Cell lysates were immunoprecipitated with anti-GFP antibodies and analyzed as described in Figure 1. (D) The interaction between recombinant GST-tagged NOT11 and an MBP fusion of NOT10 was analyzed by SDS-PAGE followed by Coomassie blue staining. The pull-downs were performed using glutathione agarose beads (lanes 4–6) or amylose resin (lanes 7–9). Input samples (1%) and bound fractions (50%) were analyzed on SDS PAGE. The asterisk indicates a contaminant protein that copurified with GST (lane 4) or bound to the amylose resin (lanes 7–9).