Fig. 6.

Detection of ubiquitinated S-RNases. (A) The pollen tube total proteins after incubation with purified S-RNase were detected with anti-S-RNase antibody by Western blot. The non-S-RNase was used as a control. The ubiquitination of S-RNase was analysed in the presence of MdSSK1, MdCUL1, S2-MdSFBB1, and His-Ub (B) or without MdSSK1 (C); reactions containing His-Ub or His-Ub and S-RNase were carried out as controls. (D) The ubiquitination of denatured S-RNase were also detected. The circles indicate the predicted size of S-RNases, the triangles indicate the ubiquitinated form of S-RNases. (E) The ubiquitination of S-RNase was analysed in the presence of MdSBP1, MdCUL1, and S2-MdSFBB1; reactions containing His-Ub or His-Ub and S-RNase were carried out as controls. By anti-MBP examination, the upper bands indicated MBP-S2-MdSFBB1, below that for MBP-MdCUL1; for anti-His, the upper and lower bands indicated His-Ub and His-S-RNase, respectively in (B), (C), (D), and (E). (This figure is available in colour at JXB online.)