Figure 2.

Expression pattern of RPBF. A, Total RNA (30 μg) from roots, seedlings, and seeds during seed development (5, 10, 15, 20, and 30 DAF) was analyzed by northern blot using gene-specific sequences downstream of the DOF domain. To compare expression patterns of the RPBF gene with GluB-1 and RISBZ1 (Onodera et al., 2001) genes, the GluB-1 coding sequence and the RISBZ1-specific sequence corresponding to the region downstream of the Leu zipper were also used as probes. 25S rRNAs are shown as a loading control. B, Total protein from embryo and endosperm of 15 DAF seeds, roots, and seedlings was analyzed by western blot with anti-RPBF, anti-RISBZ1, and anti-glutelin GluB. Protein samples were separated on a 12.5% SDS-PAGE gel and stained with Coomassie Brilliant Blue (CBB). C, Total RNAs (7.5 μg) from imbibed seeds (top section) and from deembryonated half seeds incubated in the presence or absence of GA₃ (bottom section) were subjected to northern-blot analysis. The blot was probed with the RPBF-specific sequence. Times of imbibition and GA treatment are at the top of the section.