Fig. 5

Induction of the expression of Dr-SLC38A9 upon treatment with sex-inducing substances. a-h Whole-mount in situ hybridization of Dr-SLC38A9 mRNA in sexual worms fed chicken liver (Control; a, b, e, f) or minced B. brunnea worms (c, d, g, h) daily for 3 days. Dr-SLC38A9 transcripts are stained blue. Ventral views (a-d) and dorsal views (e-h) are shown, with the anterior part of the worm at the top. b, d, f, h Higher magnification views of areas inside blue squares in panels (a, c, e, and g). White arrowheads indicate signals in the ovaries (c, d). Magenta arrowheads indicate signals in the area where supernumerary ovaries will be formed (c, d). Orange arrows indicate signals in the yolk glands (c, d). Black arrows indicate signals in the testes (g, h). i, j qRT-PCR data are shown relative to the expression level (normalized to Dr-ef1a mRNA level) in the liver-fed worm, and log₂ (relative expression) on the vertical axis indicates −∆∆Ct. The bars in the plots indicate the averages of −∆∆Ct. Asterisks indicate significant differences compared with liver-fed worms (Tukey’s HSD test: *P < 0.05; ***P < 0.001; n.s., not significant). i Six circles indicate individual samples of liver-fed worms, worms fed B. brunnea, and worms fed liver supplemented with the M0 + M10 fraction. j Twelve circles indicate individual samples of liver-fed worms and worms fed liver supplemented with l-Trp or d-Trp