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. 1997 Mar 4;94(5):1721–1726. doi: 10.1073/pnas.94.5.1721

Figure 2.

Figure 2

Stabilization of Gal80p/HA-Gal3p complex in the presence of both galactose and ATP. Yeast cells (NFG1) overexpressing Gal80p and HA-Gal3p were grown in ESGlyLac (lanes 1–3), in ESGlyLac supplemented with 2% glucose (lanes 4–6), or in ESGlyLac supplemented with 2% galactose (lanes 7–9). Whole-cell extracts prepared from these cells were subjected to immunoprecipitation. ATP (2 mM) and/or galactose (1 mM) were added to both binding buffer and washing buffer as indicated at top. Both binding buffer and washing buffer always contained NaCl at 100 mM. Immunoprecipitated proteins were subjected to SDS/PAGE, blotted to nitrocellulose membrane, and probed with rabbit anti-Gal80p antibody (Upper). The membrane blot was then reprobed with rabbit anti-Gal3p antibody (Lower). Molecular masses of standard markers are shown in kDa to the right (Upper).