Fig. 8

Subcellular localization of NRSYM1. a–e Immunohistochemistry of the NRSYM1-myc protein in root apical cells. A polyclonal anti-myc antibody and an antibody conjugated to Alexa Fluor 488 (green signal) were used as primary and secondary antibodies. Nuclei were visualized with DAPI (blue signal). Plants with transgenic hairy roots carrying the pLjUBQ:NRSYM1-myc construct were grown in the absence of nitrate for 3 days. N-starved plants were transferred to (a) N-free or (b) 10 mM KNO₃ medium for 20 min or (c) 24 h. d N-starved plants were transferred to 10 mM KNO₃ medium for 20 min, and then to N-free medium for 20 min. e N-starved plants were first incubated with leptomycin B (LMB) in a N-free medium for 3 h, and then transferred to 10 mM KNO₃ medium with LMB for 20 min and to N-free medium with LMB for 20 min. Scale bars: 50 μm. f The ratio of NRSYM1-accumulating nuclei. Using respective fluorescent images, the percentages of nuclei having green signals among all DAPI-stained nuclei were calculated. Error bars indicate SEM (n = 5 roots). Columns with the same lower-case letter indicate no significant difference (Tukey’s test, P < 0.05)