FIGURE 5: Bck1 is not required for Slt2 T-loop phosphorylation and cyclin C destruction following hyperactivation of the CWI pathway.

(A) Slt2 T-loop phosphorylation (upper panel) was determined by Western blot analysis of protein extracts prepared from mid-log phase cultures (WT, RSY10, bck1∆, RSY1050, and slt2∆, RSY1057) expressing functional cyclin C-YFP (pBK37) and either RHO1G19V or a vector control. The blot was stripped and re-probed for Slt2 and cyclin C protein levels (middle and lower panels respectively).

(B) As in (A) except that Slt2 and cyclin C were monitored in bck1∆ cells expressing either a vector or a plasmid harboring a constitutively active BCK1 allele (BCK1-20).

(C) As in (A) except that Slt2 phosphorylation (top panel) was also monitored in bck1∆ cells expressing a hyper-active allele of PKC1 (PKC1R398P).

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