FIGURE 7.

Comparison of the optimized dual LUC assay versus the prior art described by Liu et al. (2014). The optimized assay conditions include using pGrDL_SPb for OsSPL14 target sequence cloning, a 10:1 ratio of rice miR529b precursor culture to target plasmid culture, 3 days incubation post-agroinfiltration of N. benthamiana leaves, and dual LUC assays on extracts diluted 20-fold. The optimized assay control consists of assays where the miR529b precursor culture was replaced with a precursor construct culture that does not target OsSPL14 sequence. The Liu et al. (2014) assay conditions include using pGreen dualLUC 3′ UTR sensor plasmid for OsSPL14 target sequence cloning, a 1:1 ratio of rice miR529b precursor culture to target plasmid culture, 2 days incubation post-agroinfiltration of N. benthamiana leaves, and dual LUC assays on undiluted extracts. The Liu et al. (2014) control consists of assays where the OsSPL14 target plasmid culture was replaced with a plasmid culture that is not targeted by rice mir529b. The results are depicted as box and whisker plots, with interquartile range boxes and min/max whiskers (n = 12).