SupplementaryFigure3. TheeffectonApm4bindingandMid2localizationofaY278mutationinthe Mid2
cytoplasmic tail. (A) GST alone; GST tagged Apm4 ; His-tagged Mid2 or Mid2 Y278 were prepared as described 
and theirbinding interaction tested using pulldown assayson beadswith 20 µM Mid2.Apm4 isclearly seen on a 
GeneronNUView gel,along with a faint band for Mid2 (arrow).Mid2 wasdetected using anti-Histagantibodiesafter
westernblotting.ThisbandwasverifiedasMid2usingmassspectrometry.(B)Mid2-GFPandMid2-Y278A-GFPwere
transformed into cells otherwise lacking mid2(KAY53) or both mid2and apm4(KAY1700).Localization after1 hour
incubation with alpha factor was observed microscopically. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
SupplementaryFigure3 
Effectofpheromoneonorganization ofactin patches in wild type and apm4∆ cells. Abp1-GFPwas
transformed into KAY120 and KAY1690 cells and visualized in live cells 90 minutes after pheromone addition.