FIG. 5.

Western analysis of initiation factors in resedimented 40S fractions from the degron mutants. (A to C) Strains YAJ43 (WT), YAJ47 (tif32-td prt1-td), and YAJ46 (sui3-td) were cultured under nonpermissive conditions and analyzed as described in the legend for Fig. 3D, except that Western analysis was performed on the resedimented 40S fractions using antibodies against the proteins listed on the left of the blots. (D and E) Data from replicate experiments of the type shown in panels A to C were quantified, and the results for the tif32-td prt1-td (D) and sui3-td (E) mutants were normalized to the values measured in parallel for the WT strain and plotted as described for Fig. 3E. (F to H) Strains YAJ43 (WT) and YAJ45 (tif5-td) were analyzed identically, and the results shown in panels G and H together with data from replicate experiments were quantified and are summarized in panel F. All of the strains used for these resedimentation experiments lack the RPL11B gene in order to increase the concentration of 48S PICs (see text for further explanation).