Fig. 4
From: Feedback inhibition of AMT1 NH4+-transporters mediated by CIPK15 kinase
CIPK15 can interact with AMT1;1. Interaction growth assay (a), β-galactosidase staining (b), and filter (c) assay in yeast; and split-fluorescent protein interaction assay (d) in tobacco leaves for AMT1.1 and CIPK15 protein interactions. a Plasmids expressing AMT1;1 and CIPKs were expressed in yeast. Interaction indicated by growth on SD-Trp -Leu -His. Growth on SD-LT as control (Figure S5). Comparable results were obtained in three independent experiments. b, c Interaction of CIPKs and AMT1;1 in a split-ubiquitin system detected by X-Gal staining and filter assays using full-length AMT1;1-Cub-PLV as bait and NubG, NubI, and NubG-full-length CIPKs as prey. NubI/NubG served as positive (blue color) and negative controls, respectively. d Split-fluorescent protein interaction assay for AMT1;1 and CIPK15. YFP/chlorophyll, merged image of fluorescence and chloroplast. Reconstitution of YFP fluorescence from nYFP-AMT1;1 + CIPK15-cCFP and nYPF-AMT1;1 + cCFP (negative control). Comparable results with different combinations shown in Figure S7