Figure 5.

Methylation curtails Scd6 self-interaction a) GSTScd6 was pulled down from Scd6TAP expressing wild type and ∆HMT1 cells. Samples were analysed by western blotting with PAP antibody followed by stripping and anti-GST staining. Empty vector served as control. b) Quantitation of three independent experiments performed as in A (p=0.029). c) Localization of Scd6GFP to foci was checked in SCD6GFP and SCD6GFP∆HMT1 strains expressing Scd6mCherry. Cells were grown until OD₆₀₀ 0.35–0.5 followed by live cell imaging. d) Quantitation of three independent experiments performed as in c (p=0.045). e) Recombinant purified in vitro methylated and unmethylated Scd6FLAG was incubated with GST and GSTScd6 followed by Glutathione pull down and western blotting with anti-GST and anti-FLAG antibody to check their interaction (A lane has been omitted in the blots between second and third lane). Cross-reactivity to GST antibody observed in lanes 2 and 4 was due to breakdown of GSTScd6. f) Quantitation of three independent experiments performed as in e (p=0.0017).