EcR-USP RNAi resulted in significant death phenotypes and disrupted 20E signaling. (A) Typical prepupal death or pupal death phenotypes. (B) The ratio of prepupal death or pupal death. (C-D) USP mRNA (C) and USP protein (D) were decreased by USP RNAi. (E-F) mRNA levels of the two 20E primary-response genes E75B (E) and Br-C (F) were decreased. Black: control; gray: experimental. Thirty animals were used for each group and 3 biological replicates were conducted in the RNAi experiments. *: 0.01 < p < 0.05; **: p < 0.01.