FIGURE 3.

BrZ7 silencing inhibited the metamorphosis and 20E response gene expression. The 6th-72 h larvae were injected with 1 μg of dsBrZ7. The control larvae received the equivalent volume of dsGFP. Sixty larvae were used in every treatment, and the experiments were conducted in triplicate with independent experimental samples. A, phenotypes after dsGFP or dsBrZ7 injection. The black square shows that BrZ7 was knocked down in the fat body of 6th-96 h, as analyzed by Western blot with 7.5% SDS-PAGE. β-Actin was used as an internal control treatment. Scale bar, 1 cm. B, percentage of different phenotypes after dsBrZ7 was injected. C, relative expression of the 20E-induced genes analyzed by qRT-PCR. RNA was isolated from the fat body of the 6th-96 h larvae. The results were based on the 2^−ΔΔCT method in the presence of normalized β-actin gene. Asterisks, significant differences (Student's t test; *, p < 0.05).