Figure 1.

Weak association of Gal80p with HA-Gal3p in the absence of galactose and ATP. Yeast cells (NFG1) overexpressing Gal80p and HA-Gal3p were grown in ESGlyLac supplemented with glucose (lanes 1–3) or galactose (lanes 4–6) at 2%. Whole-cell extracts from these cells were incubated with 12CA5 anti-HA antibody conjugated with protein G/A agarose. NaCl was added to both binding buffer and washing buffer at the indicated concentrations. Immunoprecipitated proteins were subjected to SDS/PAGE, blotted to nitrocellulose membrane, and probed with rabbit anti-Gal80p antibody (Upper) or rabbit anti-Gal3p antibody (Lower).