Fig. 6.

Ctr9/Paf1 subcomplex formation is essential for yeast Paf1C assembly. a, b Co-IP experiments of Paf1C formation by Ctr9 (a) and Paf1 (b). Extracts were prepared from HEK293T cells transfected with various combinations of plasmids, as indicated, immunoprecipitated with agarose-conjugated anti-GFP and subsequently immunoblotted with anti-Myc or anti-GFP, as indicated. The top panel shows the IP results. The middle panel represents the IP of GFP and GFP fusion proteins (GFP-Ctr9 or GFP-Paf1). The bottom panel shows 3% of the Myc fusion proteins for each IP. The asterisk indicate the degradation of Myc-Ctr9. c Model of yeast Paf1C assembly. The Ctr9/Paf1 heterodimer is the core component for Paf1C assembly. The bold line represents the interaction in the crystal structure of the Ctr9^(1–313)–Paf1^(34–103) heterodimer or the interaction between Paf1 and Leo1 obtained from the IP results and studies^45,46. The fine lines represent the interaction obtained from the IP results. The dotted lines represent interactions that need to be further studied. Uncropped blots are shown in Supplementary Fig. 8