Fig 1. Scd6 interacts with Hmt1.

(A) Two-hybrid analysis; The yeast strain PJ69-4A was co-transformed with pGBD-c1 or pGBD-c1-SCD6 with either pGAD-c1 or pGAD-c1-HMT1 and colonies were spotted onto SC-Leu-Trp (WL) and SC-Leu-Trp-His (WLH) plates and incubated for 3 days at 30°C. (B) Co-immunoprecipitation analysis; Scd6Flag was immunoprecipitated from Hmt1myc strains harboring YCplac33-SCD6FLAG or empty vector with an anti-Flag antibody, and were probed for Scd6Flag and Hmt1myc using anti-Flag and anti-myc antibodies, respectively. Similar experiments were performed using total extracts treated with RNase A prior to immunoprecipitation. (C) In vitro binding assays; His₆-Hmt1 was incubated with Glutathione sepharose beads that had been preloaded with equal amounts of GST or GST-Scd6. Recombinant proteins were then detected using Coomassie staining and associated His₆-Hmt1 was detected using an anti-His antibody.