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. 2000 Sep;11(9):2873–2884. doi: 10.1091/mbc.11.9.2873

Figure 3.

Figure 3

Binding-defective receptors, Ste2-S184R, undergo ligand-induced endocytosis when expressed with wild-type receptors. Each of the four strains analyzed expressed an HA-tagged receptor (Ste2-HA or Ste2-S184R-HA) encoded by a chromosomal allele and an untagged plasmid-encoded receptor (Ste2 or Ste2-S184R). Log-phase cultures in −Ura + CAA medium were treated with cycloheximide and then cultured for 15 min in the absence (○) or presence of α-factor (●). Membranes were fractionated by using Renografin density gradients. Fractions were assayed for HA-tagged receptors (○,●) and for plasma membrane ATPase (no plot symbol) by using immunoblotting methods. Plasma membrane marker is shown for α-factor-treated cultures only. Protein amount is the percentage of the total protein. (A) Cells expressing Ste2-HA and Ste2 (strain DJ1419-A). (B) Cells expressing Ste2-S184R-HA and Ste2-S184R (strain DJ1418-A). (C) Cells expressing Ste2-HA and Ste2-S184R (strain DJ1403-A). (D) Cells expressing Ste2-S184R-HA and Ste2 (strain DJ1402-A).