Effects of AeELO2 and AeELO9 knockdown on the growth rate and cold tolerance of larvae and pupae. (A) Larvae to pupae development rate. The days after treatment and the percentage of pupae are shown on the x- and y-axes, respectively. Each treatment group contained 120 larvae, and the experiment was repeated three times. (B) Mortality rate of the 4th instar larvae at 4 °C. The days after treatment and mortality rate (%) are shown on the x- and y-axes, respectively. (C) Mortality rate of pupae at 4 °C. The days after treatment and mortality rate (%) are shown on the x- and y-axes, respectively. (D) The osmotic balance of larval cuticle. The larvae of different treatment groups were soaked with Eosin Y dye for 5 min. Here, 1 represents the larvae of blank control; 2 represents the larvae treated with gus-dsRNA; 3 represents the larvae in the AeELO2-dsRNA treatment group; 4 represents the larvae treated with AeELO2-dsRNA. Each treatment group contained 30 larvae, and the experiment was repeated three times. Abbreviations: DEPC, blank control, larvae treated with diethyl pyrocarbonate water; GUS, larvae treated with gus-dsRNA; ELO2, larvae treated with AeELO2-dsRNA; ELO9, larvae treated with AeELO9-dsRNA; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant. *, **, etc., represent a significant difference compared with the blank control (DEPC water-treated).