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Fig. S5 Reduction of wnt8b affects emx2 expression in the posterior-ventral hypothalamus. (A-I) WISH of 1-dpf wild-type embryos (A,D,G) and embryos injected with 3 ng wnt8b MO (B,E,H) or 2.5 ng fgf8 MO (C,F,I) with probes for emx2, nkx2.1a and shh shows that loss of wnt8b affects emx2 expression. (A-C) emx2 expression is reduced in the posterior-ventral hypothalamus in wnt8b morphants (B, arrowhead; n=19/19) but is not changed in fgf8 morphants (C; n=18/18). (D-F) Neither wnt8b (E; n=17/17) nor fgf8 (F; n=19/19) morphants show changes in nkx2.1a expression. Also, the expression of shh is unchanged upon loss of wnt8b (H; n=24/24) or fgf8 (I; n=18/18). (J) One-dpf wild-type embryos subjected to two-color FISH to analyze the relative hypothalamic domain sizes of emx2 and nkx2.1a using ImageJ. (K) The ratio of the nkx2.1a expression domain to the hypothalamic emx2 expression domain is unchanged in fgf8 morphants. The number of analyzed embryos is indicated within the bars. P=0.2483 (Student’s t-test); n.s., not significant. Lateral views are shown. Tel, telencephalon; Hyp, hypothalamus; LOF, loss-of-function. Scale bars: 100 μm in A-I; 60 μm in J.