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Fig. 5. MMS-induced Chk1 phosphorylation is abrogated in rad9{Delta}1-231 mutants. Indicated strains containing the cdc15-2 allele were transformed with a CHK1-HA plasmid and cultured in selective media. Cultures were arrested in telophase by incubation for 3 hours at 37°C before being mock treated or treated with 0.1% MMS (v/v) for 1 hour. Whole cell extracts were prepared and immunoblotted with {alpha}-Rad9, {alpha}-Rad53 or {alpha}-HA antibodies. The same phosphorylation patterns were observed with two separate extracts and Chk1 phosphorylation was detected with both HA- and Myc-tagged proteins.





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