Figure 9

APUM5 negatively regulates the 3′ UTRs of ABA-responsive genes in the reporter assay. (A and B) Protoplasts of Col-0 and 35S-APUM5 transgenic plants were transformed with the RD22-3′ UTR or RAB18-3′ UTR reporter construct by PEG-mediated transformation. GFP signals were detected by LSM 700 confocal microscopy under identical conditions. (C, D) GFP signal intensity of the RD22-3′ UTR and RAB18-3′ UTR reporter were analyzed with Zen software of the LSM 700 confocal microscope and ImageJ software (rsbweb.nih.gov/ij/). Error bars represent ± SD (Student’s t-test; **P < 0.001). (E, F) Western blot and RT-PCR analysis were performed with protoplasts of Col-0 and 35S-APUM5 transgenic plants transformed with the RD22-3′ UTR or RAB18-3′ UTR reporter construct. Protein and RNA samples were obtained from four-independent experiments and extracted. Rubisco was used as the protein loading control and AtActin7 was used as the internal control. The numbers designate relative intensity.