CsSCLs were the targets of csi-miR171c and CsSCL proteins located in cell nucleus. A) The sequences of native and mutant MRE of csi-miR171c on CsSCL (CsSCL-nMRE and CsSCL-mMRE). The double, single, and no point represent perfect, imperfect, and no complementation, respectively. The red nucleotides were modified. B) Diagrams of the overexpression vectors constructed for transient expression in N. benthamiana. oe, Pre-miR171c overexpression vector; ev, empty vector; n, the overexpression vectors of the native MRE with C-terminal fusion protein; m, the overexpression vectors of the mutant MRE with C-terminal fusion protein. C) GFP fluorescence on the N. benthamiana leaves injected with Agrobacterium tumefaciens. Photos were taken under bright field and UV light (excitation light wavelength 365 nm) 3 d after infection. WT leaf without any treatment showed no fluorescence signal under UV. Scale bars = 1 cm. D) The fluorescence readout ratios between different coexpression combinations within the injection areas of N. benthamiana leaves. Results are means of fluorescence readout ratio ± SEM from 4 leaves. Asterisks indicate statistically significant differences of CsSCL-nMRE compared with CsSCL-mMRE using an independent 2-sample t-test (*P < 0.05, **P < 0.01). E) Cleavage sites of csi-miR171c at MRE in CsSCLs (CsSCL1, CsSCL2, and CsSCL3) determined by 5′-RACE analysis. The arrows indicate the cleavage sites at MRE, and the numbers indicate the frequency of valid sequenced clones. F) Subcellular localization of CsSCL1, CsSCL2, and CsSCL3. CsSCL1-YFP, CsSCL2-YFP, CsSCL3-YFP, and empty vector-YFP, respectively, were cotransformed with OsGhd7-CFP (a nuclear marker) into Arabidopsis mesophyll protoplast. The photographs were captured with filters for YFP (excitation 514 nm/emission 550 to 590 nm), CFP (excitation 448 nm/emission 460 to 505 nm), bright light, and merged. Scale bars = 10 μm.