Fig. 2

Developmental expression and JH induction of HDAC3 in T. castaneum. A HDAC3 mRNA levels were determined during the penultimate, last larval, and pupal stages at 24 h intervals. Total RNA isolated from a pool of two larvae for each replication was converted to cDNA and used in RT-qPCR to determine the relative HDAC3 mRNA levels. Mean ± SE (n = 4) are shown. Levels not connected by the same letter are significantly different. B JH suppresses the expression of HDAC3 in T. castaneum larvae. S-Hydroprene (H, JH analog) was dissolved in cyclohexane (C) and topically applied to 48 h-old last instar larvae (0.5 μL of 2 μg/μL). At six hours after treatment, total RNA was isolated and subjected to RT- qPCR. The expression of the JH response gene Kr-h1 was significantly induced, and HDAC3 was significantly suppressed, mean ± SE (n = 4) are shown. Levels not connected by the same letter are significantly different. C Met is required for suppression of HDAC3 by hydroprene. Newly molted last instar larvae were injected with dsMet or dsmalE. At 48 h after injection of dsRNA, the larvae were treated with hydroprene. Total RNA isolated from larvae was converted to cDNA and used to quantify Kr-h1, HDAC3 and, Met mRNA levels. The data shown are mean ± SE (n = 4). The data were analyzed using analysis of variance, each pair student’s t-test. Mean values with the same letter are not significantly different from each other. C, cyclohexane; H, hydroprene