Figure 4.

AmiRNA to CKB delays circadian period in Arabidopsis protoplasts. A, The average bioluminescent readout (CCA1-LUC) from protoplasts transfected with amiR-CKBfam. Experiments were performed as described in Figure 1. Data represent mean values ± SEM (n = 6). Similar results were obtained from two independent trials. B, The period length (CCA1-LUC) from protoplasts transfected with combinatorial sets of amiRNA targeting each CKB subunit. Experiments were performed as described in Figure 1 except for using different amounts of plasmid for each combination (one, two, three, and four effector plasmids: 55 μg, 22.5 μg, 11.3 μg, and 5.7 μg per each plasmid, respectively). Data represent mean ± 95% confidence interval (n = 6). Similar results were obtained in two independent trials. C, Relative expression of four CKB subunit genes from cDNA of protoplasts transfected with control amiRNA, or amiR-CKB1, -CKB2, -CKB3, or -CKB4. Protoplast harvest and determination of gene expression was performed as described in Figure 3B. Data represent mean ± SEM (n = 3). D, Relative expression of four CKB subunit genes from cDNA of protoplasts transfected with control amiRNA or amiR-CKBfam. Protoplast harvest and determination of gene expression was performed as described in Figure 3B. Data represent mean ± SEM (n = 3).