Skip to main content
. 2018 Mar 15;29(6):736–750. doi: 10.1091/mbc.E17-08-0518

FIGURE 4:

FIGURE 4:

Exocyst subunits with a carboxy terminal TM tail were largely absent from the 50,000 × g supernatant. (A) Total cell lysates (T) from WT strains expressing 3xHA or 2xHA-sso1TM at the C-terminal end of EXO70, SEC3, or SEC6 (NY3241, NY3252, NY3242, NY3253, NY3251, and NY3255) were centrifuged at 50,000 × g, separated into supernatant (S) and pellet (P) fractions, and analyzed by Western blot analysis on an 8% SDS–PAGE gel. Tagged proteins were detected with anti-HA antibody. Mutant strains are heterozygous diploids with one intact WT copy. (B) Total cell lysates (T), 50,000 × g supernatant (S) and pellet (P) fractions from heterozygous diploid strains, SEC10-2xHA-sso1TM/WT (NY3257) or SEC15-2xHA-sso1TM/WT (NY3258), were analyzed by Western blot with anti-Sec10p or anti-Sec15p serum. Total lysate from WT (NY1210) was used as a control. (C) Total cell lysates (T), 50,000 × g supernatant (S), and pellet (P) fractions from pEXO70-EXO70-3xHA (NY3241) or pADH-exo70-2xHA-sso1TM (NY3268) were analyzed by Western blot with anti-HA antibody. Control membrane protein Sso1/2p was detected with anti-Sso1/2 and Snc1/2p was detected with anti-Snc1/2 sera.