Pleiotrophic effects of SBEI and SBEIIb down-regulation on other starch biosynthesis-related enzymes in developing endosperm at 10 DAF in 2016. A, Reverse transcription-PCR analysis of starch biosynthesis-related enzyme genes. The actin gene is used as a reference for comparative quantitation. Values are means ± sd from three biological replicates each with technical triplicates. Asterisks highlight significant differences in starch synthesis-related enzymes between TQ and TRS by Student’s t test (***, P < 0.001). B, Immunoblotting of starch biosynthesis-related enzymes. Anti-HSP82 antibody is used as a loading control. C, Native PAGE/activity staining of SBEs, SSs, Pho, and DBEs. D, GBSSI activities of starch granules isolated from TQ and TRS endosperm. Values are means ± sd from three biological replicates.