Skip to main content
. 2001 Feb;12(2):421–435. doi: 10.1091/mbc.12.2.421

Figure 3.

Figure 3

Rsp5p associates with the membrane fraction of a cell lysate. (A) Differential centrifugation of a lysate prepared from cells expressing HA-Rsp5p (LHY2066). LHY2066 cells were propagated at 24°C in YPUA galactose medium; 13,000 × g (P13), 100,000 × g (P100), and soluble (S) cell fractions were resolved by SDS-PAGE, transferred to nitrocellulose, and probed with HA antibodies. (B) Differential centrifugation after biochemical treatments. LHY896 cells were grown in YPUAD medium at 30°C. The cell lysate was mock (buffer) treated, treated with 2.5 M urea, or treated with 1.0% (vol/vol) Triton X-100 (TX100) and 0.2% (wt/vol) dodecyl β-d-maltoside (β-DM) for 1 h on ice before fractionation. Fractions were analyzed as in A by immunoblotting with HA, c-myc, or hexokinase antibodies to detect HA-Rsp5p, myc-Emp47p, and hexokinase, respectively. An equivalent amount of the total cell extract is shown for comparison. SUP, supernatant.