Figure 2.

Ubiquitin Binding Activity and Gene Expression Analysis of OsDSK2a.

(A) Binding of OsDSK2a to poly-ubiquitin (poly-Ub) chains in vitro. The OsDSK2a-GST fusion protein was precipitated by glutathione beads and incubated with a mixture of poly-Ub chains linked via K48 or K63. The flow-through was collected (Flow), and the beads were washed three times (W1–W3). The precipitated proteins were eluted (Elute) and separated by SDS-PAGE before immunoblotting with anti-Ub antibodies.

(B) pOsDSK2a: GUS expression patterns in transgenic rice plants at the seedling stage. Bars = 1 cm.

(C) pOsDSK2a: GUS expression patterns in transgenic rice plants at the heading stage. (1) to (5) Nodes from top to bottom. (6) Last internode and root. (7) Elongated zone of the uppermost internode. (8) Elongating zone of the uppermost internode. (9) Division zone of the uppermost internode. (10) to (13) Second to fifth internodes. (14) Young spikelets. (15) Flag leaf. (16) Anthers. Bars = 1 cm.

(D) qPCR analysis of OsDSK2a expression in different tissues. ACTIN was used as a control. The data represent the means ± sd (n = 3).