Fig. S4.
The CC domains of MLA10 and Sr50 do not self-associate in yeast. (A) The yeast strain HF7c was cotransformed with the specified constructs, and self-interaction of MLA10 and Sr50 CC domain fragments (BD:MLA101–160/AD:MLA101–160, BD:MLA101–225/AD:MLA101–225, BD:Sr501–163/AD:Sr501–163, BD:Sr501–228/Ad:Sr501–228, and BD:Sr501–128/AD:Sr501–128) was assayed by yeast two-hybrid assay. AD:RGA51–228, BD:RGA51–228, AD:L6-TIR, and BD:L6-TIR constructs were used as controls. Cultures of cotransformed yeast clones were adjusted to an OD of 0.2, and three dilutions (1/1, 1/10, 1/100) were spotted on synthetic −LTH medium (−Trp/−Leu/−His) to assay for interactions and on synthetic double-dropout −LT medium (−Trp/−Leu) to monitor proper growth. Photographs were taken after 4 d of growth. (B) Myc- or HA-tagged proteins were extracted from yeasts and detected by immunoblotting using anti-Myc or anti-HA antibodies, respectively.