Detection of the interaction between IAA8 and ARFs by yeast two-hybrid and BiFC assays.

<p>(A) Yeast (EGY48::pJK103) cells were co-transformed with BD-IAA8 bait plasmid and AD-ARFCTD prey plasmid as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0043414#pone-0043414-g004" target="_blank">Figure 4</a>. The strength of each interaction was evaluated by measuring β-gal activity using the o-nitrophenyl β-D-galactopyranoside (ONPG) method. Vector indicates empty vector (negative control experiment). (B) A BiFC assay was carried out to monitor the protein-protein interaction between IAA8 and ARFs. <i>ARF-nYFP</i> fusions or <i>nYFP-GUS</i> and <i>IAA8-cYFP</i> were transiently co-expressed in <i>Arabidopsis</i> mesophyll protoplasts with a nuclear marker plasmid, NLS-tdTomato. BiFC, NLS, and BF (top) represent YFP, tdTomato fluorescence, and bright field images, respectively. n: nucleus. Bars = 10 μm.