Figure 4. BIN2 interacts with EGL3, TTG1, and WER.
(A) BIN2 interacts with EGL3 in yeast two-hybrid assays. (B) The interaction of BIN2-His with CPC-GST, EGL3-GST, TTG1-GST, and WER-GST in vitro. The BIN2-His pulled-down by CPC-GST, EGL3-GST, TTG1-GST, and WER-GST, or GST was detected by western blotting with anti-His antibody (top). The purified BIN2-His protein was used as inputs. An equal loading of recombinant proteins was indicated by Coomassie brilliant blue (CBB) staining (bottom). (C) BiFC assays of the interaction between BIN2 with EGL3, TTG1, and WER. Scale bars, 20 μm.
Figure 4—figure supplement 1. Yeast two-hybrid assays to test interactions of BIN2 with WER, TTG1, or CPC.
The full-length cDNA of each corresponding gene was fused with the GAL4-BD or AD domain. Yeast cells harboring the indicated constructs were grown on the synthetic media lacking Trp and Leu, or Trp, Leu, and His with an additional 1 mM 3AT.