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. Author manuscript; available in PMC: 2010 Nov 7.
Published in final edited form as: Traffic. 2006 May;7(5):574–588. doi: 10.1111/j.1600-0854.2006.00410.x

Figure 1. Clathrin LC is important for Sla2p–HC interaction, but clathrin HC is not required for Sla2p-LC binding.

Figure 1

(A) Wild-type (WT) (YPJ96-4), clc1Δ (SL4410) and chc1Δ (SL4409) two-hybrid reporter strains were transformed with GAL4-binding domain (GBD) bait plasmids pGBDU (GBD empty), pKH19 (GBD-CLC1) or pKH47 (GBD-sla2-292-501) in combination with GAL4 activation domain (GAD) prey plasmids pGAD (GAD empty), pKH24 (GAD-chc1-655-1653) or p31-2 (GAD-sla2-292-520). Equal numbers of cells from transformants were spotted on complete synthetic medium lacking leucine and uracil (C-LEU-URA) and medium lacking adenine, leucine and uracil (C-ADE-LEU-URA) and grown for 5 days to score for ADE2 reporter expression. (B) Bacterially purified GST fusions prebound to glutathione sepharose beads were incubated with yeast lysates from WT (SL1629) or clc1Δ (SL1628) to pull down clathrin HC. GST-Bmh1p was used as a control for nonspecific binding. (C) Same as (B), except GST fusions were incubated with yeast lysates from WT (SL1463 + pKH2) or chc1Δ (SL1927) to pull down clathrin LC. (B and C) Blots were probed with indicated antibodies. Ten microliters of cell lysates were run as input controls (lanes 1 and 6).