Figure 5.

Effects of OsRac1-RNAi and heterotrimeric G-protein Gα mutation on OsMAPK6 protein, gene expression, and kinase activity. A, Total soluble proteins were extracted from cv Kinmaze (wild-type) cells and from cell lines with a silenced OsRac1 gene (OsRac1-RNAi) and analyzed by immunoblotting with the anti-OsRac1 antibody (top), anti-OsMAPK6 antibody (middle), or anti-α-tubulin antibody (bottom) as the loading control. B, Total RNAs were isolated from OsRac1-RNAi lines and analyzed by RT-PCR using specific primers of OsMAPK6 and ubiquitin. C, The OsRac1-RNAi line (line 1) and d1 mutant were treated with elicitor 5 μg mL⁻¹ SE, sampled at the indicated times, and assayed for in-gel kinase activity. D, Total RNAs were isolated from wild-type cells and the heterotrimeric protein Gα-subunit null mutant (d1) and analyzed by RNA gel blot. Membranes were probed with a [³²P]-labeled OsMAPK6 gene-specific probe. E, Total soluble proteins were extracted from wild-type cells and the heterotrimeric protein Gα-subunit null mutant (d1) and analyzed by immunoblotting with the anti-OsMAPK6 antibody and anti-α-tubulin antibody as the loading control.