Fig. 3. The N domain of Ski7p interacts with the components of the Ski complex and the exosome. Cell extracts were obtained from the indicated yeast strains expressing Ski2p-Myc and Ski8p-HA (A–E) or Prp4p-Myc and Ski6p-HA (F–J). The yeast strains also expressed the indicated forms of Flag-tagged Ski7p (A–E, lanes 1–4 for MC146–MC149; F–J, lanes 1–4 for MC151–MC154). The cell extracts were immunoprecipitated (IP) with an anti-Flag antibody. The precipitated proteins (B, D, E, G, I and J), together with the whole-cell extracts (A, C, F and H), were immunoblotted (IB) with anti-Myc (9E10), anti-HA (12CA5), and anti-Flag (M2) antibodies to detect Ski2p (or Rrp4p), Ski8p (or Ski6p) and Ski7p. The asterisks in (E) and (J) indicate Flag-tagged Ski7p mutants expressed in the various strains. The arrowheads indicate the positions of IgG bands.