Fig. 3. Wnt3a induces Irx3 and Gbx2 in prospective dT explants. (A) The experimental procedure. Prospective vT and dT explants from HH stage 8 embryos were embedded in collagen, cultured in the presence or absence of soluble Wnt3a, fixed and assayed for Irx3, Gbx2 and Dlx2 expression by in situ hybridization. (B) The data was quantified and analyzed statistically using the -square significance test. (C-N) Representative explants in the culture condition indicated following in situ hybridization for Irx3 (C-F), Gbx2 (G-J) and Dlx2 (K-N). Some panels show more than one explant. The total number of explants in each experimental condition are indicated above each bar in B. *** indicates P<0.0001. Scale bar: 250 µm in C-F, 100 µm in G-N.