Drawing of the BiFC assay. The estradiol-inducible C-terminal yellow fluorescent protein (YFP) fragment Vc is fused to Stm1 (orange), and the constitutively expressed N-terminal YFP fragment Vn is cloned in frame with Rpl42a. Interaction of Stm1 with Rpl42a or Rrp7 reconstitutes YFP fluorescence (top). Representative images of estradiol-induced (10 nM, 2 h) cells expressing Vc-Stm1 (Stm1) or Rpl42a-Vn (Rpl42a) or Vc-Stm1 and Rpl24a-Vn (Stm1 + Rpl42a) are shown (middle). Scale bar: 5 μm. Merged YFP and bright-field images are shown (middle). BiFC quantification is shown as percentage of YFP positive cells (bottom graph) from at least 50 cells/sample and independent replicate (dots) (n =3). Data values are shown as mean and standard deviation.
