FIG. 1.

Kinetics of Rvb2 depletion in galactose-dependent test cells. Test cells dependent on a GAL::rvb2 allele (strain YWD313) were grown in medium containing 2% galactose, washed, and shifted to medium containing 2% glucose. Cell samples were collected before and at various intervals after the medium shift. (A) Northern analysis of RVB2 mRNA accumulation. Total RNA was extracted from cells at the time points indicated and fractionated on an agarose-formaldehyde gel, and a blot was probed with radiolabeled DNA that recognizes RVB2 mRNA. (B) Western analysis of Rvb2p protein accumulation. Total protein was extracted at the time points indicated, fractionated by SDS-polyacrylamide gel electrophoresis, and electroblotted to a nitrocellulose membrane. Proteins were detected with rabbit polyclonal IgG (raised against recombinant Rvb2p) and a goat anti-rabbit–peroxidase conjugate. Left lane, total protein from a strain (YWD312) carrying HIS3::rvb2 and Rvb2-GFP expressed from a plasmid; the other lanes show total proteins from GAL::rvb2 cells incubated in glucose for 0, 2, and 8 h, as indicated.