Figure 9.
The intracellular distribution of Lst1p. (A) Cells expressing Lst1p-HA from a centromeric plasmid (CKY535) were gently lysed and subjected to sequential centrifugation steps, giving 500 g, 10,000 g, and 150,000 g pellet fractions (P) and a 150,000 g supernatant fraction (S). Each sample contains extract from the same number of cells. (B) Cell lysates were treated for 1 h at 4°C with either 2.5 M urea, 500 mM NaCl, 100 mM sodium carbonate (pH 11.5), or 1% Triton X-100. Pellet (P) and supernatant (S) fractions were then separated by centrifugation at 50,000 g. Lst1p-HA was detected by SDS-PAGE and immunoblotting with anti-HA antibody.