Fig. 10.

WT and mutant Dre2 expression levels are comparable. WT (BY4742) cells were grown in either CSM or YPD medium. Dre2-TAP cells carrying various Dre2 alleles on pRS415 plasmid and empty pRS415 plasmid as control were grown in CSM-Leu medium. Equivalent cell numbers (4 × 10⁶) were loaded in each lane of a 12% SDS–PAGE gel and proteins were electrophoretically separated. Proteins were then transferred to a nitrocellulose membrane for immunoblotting using polyclonal anti-Dre2 antibody. (A) Amido black staining of the membrane was photographed to show equal protein loading. (B) Statistical comparison of Dre2 levels in different cells. (C) Triplicate Western blots were performed. In each blot, Dre2 levels were normalized to the protein levels in the bracketed area using Image J software. Dre2 level in WT cells grown in CSM medium was set to 1.