Fig. 5.

In vivo and in vitro assay of OsLFL1 binding to the promoter of OsMFT1. (A) The promoter of OsMFT1 was divided into four fragments (pro1–4), and the RY motif was contained in pro4. (B) OsLFL1 bound to pro4 in yeast cells through a yeast one-hybrid assay on selective medium (SD/−Trp−Ura) containing X-gal for developing the blue color. (C) EMSA assay using the OsLFL1 protein and 50-bp OsMFT1 promoter containing RY motif as a probe labeled with 5′-biotin. The 10-, 20-, 50- and 100-fold non-labeled probes were used for competition. (D, E) OsLFL1 activates the expression of OsMFT1 by dual luciferase transient assay in rice protoplasts. Error bars indicate SD based on three biological replicates, **P<0.01, t-test.