ilp2, -6, -4, and -5 are affected by the Aa5HT2B CRISPR-Cas9 depletion. (A) Expression levels of ilp2 and ilp6 transcripts are significantly lower, but those of ilp4 and ilp5 are significantly higher in ΔAa5HT2B mutant adults (T1 and T2 females at 24-h PBM with reduced body size) than WT controls. The expression levels of ilp1, -3, -7, and -8 and insulin/IGF pathway components are not affected by the depletion of Aa5HT2B. (B) The ΔAa5HT2B mutant larvae (T1 and T2 larvae on the fifth day after egg hatching with reduced body size) display the same trend of regulation as adults in transcription. ilp8 is undetectable in late larvae of WT or mutant females. Ilp1, -3, -4, -7, and 8 (head-specific expression) were determined in the head; ilp6 was determined in the fat-body; other genes were determined in the whole body. Data represent three biological replicates (10 individuals in each replication) with three technical replicates and are shown as mean ± SEM. **P < 0.01; ***P < 0.001.