RTKs were screened to determine their involvement in the JH signal pathway in HaEpi cells and larvae.
(A) The roles of RTKs in JH III-induced Kr-h1, Vg, Jhi-1, and Jhi-26 expression were determined by RNAi of Rtk genes (1 μg/mL dsRNA, 48 h, 1 μM JH III for 12 h). DMSO as solvent control. The relative mRNA levels were calculated via the 2-ΔΔCT method and the bars indicate the mean ± SD. n = 3. Multiple sets of data were compared by analysis of variance (ANOVA). The different lowercase letters show significant differences. (B) The examples of phenotype after Vegfr1, Drl, Cad96ca, Nrk, Fgfr1, and Wsck knockdown in larvae. Scale = 1 cm. (C) Phenotype percentage and pupation time after Vegfr1, Drl, Cad96ca, Nrk, Fgfr1, and Wsck knockdown in larvae. The time was recorded from the bursting of the head shell of the 5th instar to pupal development. Images were collected after more than 80% of the larvae had pupated in the DMSO control group. Two-group significant differences were calculated using Student’s t test (*p<0.05, **p<0.01 indicate the significant difference between the percentages of the delayed pupation in dsGFP + JH III control group and gene knockdown) based on three replicates, n = 30 × 3 larvae.