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. 2010 Nov 22;31(3):584–598. doi: 10.1128/MCB.00845-10

FIG. 5.

FIG. 5.

Thr290 is required for Ash1 instability in vivo. (A) Wild-type strains transformed with either GAL1-ASH1HA or GAL1-ASH1HAT290A plasmids were arrested by nocodazole and subjected to promoter induction/repression for the indicated times, and Ash1HA or Ash1HAT290A was detected by anti-HA immunoblotting and quantified. Diamonds indicate negative-control samples not induced with galactose. (B) The same experiment as in panel A was performed with α-factor-arrested strains. (C) The same experiment as in panel A was performed, except with nocodazole-arrested cdc4-1 strains and with induction/repression at 37°C. (D) The same experiment as in C was performed with α-factor-arrested strains. (E) ash1::ASH1MYC cdc15-2 and ash1::ASH1MYCT290A cdc15-2 were arrested at 37°C (t = 0) and released at 23°C for the indicated times, and Ash1MYC or Ash1MYCT290A was detected by anti-MYC immunoblotting. Ash1MYC is displayed in “+” lanes, and Ash1MYCT290A is displayed in “−” lanes. The budding index at each time point is indicated.