Fig. 5

CHIQ1 family interacts with EMF2. a Phylogenetic tree of Arabidopsis CHIQ1 family (left) made using Phylogeny.fr [121] and motif conservation in CHIQ1 protein family (right) predicted by MEME [122]. Motifs 1 and 4 correspond to the DUF641 domain. Height of the domains indicates the degree of conservation, where taller domains are more conserved than shorter ones. CHIQ1 is in blue and CHIQ1’s interactors in red. CHIQL6 (TAIR: AT1G29300), CHIQL7 (TAIR: AT2G32130), CHIQL4 (TAIR: AT3G14870), CHIQL5 (TAIR: AT1G53380), CHIQL8 (TAIR: AT2G30380), CHIQL3 (TAIR: AT4G36100), CHIQL2 (TAIR: AT4G33320), CHIQ1 (TAIR: AT2G45260), CHIQL1 (TAIR: AT4G34080), CHIQL9 (TAIR: AT3G60680), CHIQL10 (TAIR: AT5G58960). b-d Physical interaction between CHIQ1, CHIQL6, CHIQL5, and EMF2 based on yeast two-hybrid assays (b), pull-down assays in tobacco (c), and bimolecular fluorescence complementation assays in Arabidopsis protoplasts (d). Pull-down assays were performed with anti-FLAG antibody and we used anti-GFP antibody to detect CHIQ1, anti-GST antibody to detect EMF2, and anti-FLAG antibody to detect CHIQL6, CHIQL5, and EMF2 in the eluted immuno-precipitate. The input corresponds to the total protein extract and IP is the eluted immuno-precipitate. Error bars in (b) represent standard error. * = p-value <0.001, t-test. In (d), green indicates fluorescence from reconstituted Venus fluorescent protein. Red indicates autofluorescence from the chloroplast. The percentage corresponds to the fraction of cells expressing Venus in each sample. Representative images from three independent experiments are shown (n = 258–321 cells per pair per experiment)