Fig. 3

Egf ligands Vn and Spi promote JH biosynthesis. A qPCR showing the tissue-specific expression of spi, vn, and krn on 5-day-old adult females. Ov, ovary; CA, corpora allata; Mg, midgut; Cg, colleterial glands; Epi, epidermis; Fb, fat body; Br, brain. B–B’ Effects of simultaneous RNAi spi and vn on the expression levels of Jhamt (B) and Cyp15A1 (B’) in the head. C–C” Effects of simultaneous RNAi spi, vn, and krn on the expression levels of Jhamt (C) and Cyp15A1 (C’) in the head as well as Kr-h1 in the fat body (C”). D–D’ Effect of simultaneous RNAi spi and vn on the JHAMT protein levels in the head (D). Quantification of band intensity of JHAMT protein level (D’). E–E’ Effect of simultaneous RNAi spi, vn, and krn on the JHAMT protein levels in the head (E). Quantification of band intensity of JHAMT protein level (E’). *P < 0.05, **P < 0.01, ***P < 0.001, n = 3. F–F” Effects of simultaneous RNAi spi, vn, and krn on the size of maturing oocytes and each follicle cell as well as follicular patency formation (F). Quantification of the length of maturing oocyte (F’) and the area of each follicle cell (F”). ***P < 0.001, n = 16 or 20. Arrow: follicular patency, blue, DNA; red, F-actin; scale bar: 1000 μm or 20 μm. G JH III titer measurements in the hemolymph. ***P < 0.001, n = 3. H–H” Effects of simultaneous spi, vn, and krn RNAi on the morphology of the CA and CA cell (H). Blue, DNA; red, F-actin. Scale bar: 300 μm or 10 μm. Quantification of the areas of CA (H’) and each CA cell (H”). **P < 0.01, ***P < 0.001, n = 20 or 21. I Effects of treatment with methoprene on the size of ovaries and follicle cells after simultaneous RNAi spi, vn, and krn. Blue, DNA; red, F-actin; scale bar: 1000 μm or 20 μm. J–K Effects of treatment with methoprene on the expression of Kr-h1 and Vg in the fat body after simultaneous RNAi spi, vn and krn. *P < 0.05, **P < 0.01, n = 3. Ace, acetone; Meth, methoprene; S, spi; V, vn; K, krn