Figure 3.
Interaction of BTS with PYEL depends on the presence of the RING (E3 ligase) domain. A, BiFC assay showing in planta interactions of ILR3, bHLH104, bHLH115, and PYE with BTS and BTSΔE3 deletion protein in leaf epidermal cells. Indicated proteins are transiently coexpressed in red fluorescent protein (RFP)-Histone2B (nuclear marker) transgenic N. benthamiana plants. Enhanced YFP (EYFP) fluorescence indicates interaction between proteins in the nucleus. All interactions were tested using both combinations of N-/C-EYFP-fused reciprocal proteins. Results shown represent three independent assays. B, Immunoblot analysis showing Co-IP of in vitro-translated Myc-tagged ILR3, bHLH104, and bHLH115 by 3xHA-tagged BTS (full length) but not by Halo-tagged BTSΔE deletion protein. Results shown represent two independent assays. Bars = 20 μm.