Fig. 3.

Unfolded protein responses in OsHrd3 KD seeds. (A) Immunoblot analysis of total proteins extracted from mature WT and OsHrd3 KD seeds. Total proteins were extracted from mature seeds with SDS–urea buffer containing 2-mercaptoethanol. The total proteins were separated by SDS–PAGE, followed by immunoblot analyses using antibodies against ER-resident chaperones. The two tracks in the WT and OsHrd3 KD represent different samples from different seeds. (B) Induction of ER stress-related genes during seed development in WT and OsHrd3 KD seeds. Total RNAs were isolated from seed tissues at 0, 7, 14, and 21 days after flowering (DAF). Transcript levels were estimated by semi-quantitative RT-PCR. 17S rRNA was analysed as a loading control. The arrowhead indicates mature transcript produced by unconventional splicing of precursor OsbZIP50 transcript.