Figure 3
From: Temporal and spatial requirements of Smoothened in ventral midbrain neuronal development
Spatial distribution of Sonic hedgehog (Shh) signaling effectors in ventral midbrain progenitor domains. (A-C) Double immunofluorescence staining of Shh and Nkx6.1 reveal that Shh proteins were expressed mainly in (A) the D1 region at embryonic day (E)9.5,(B) D1 and D2 regions at E10.5, and (C) D2 region at E11.5. (D-F) Double immunofluorescence staining of Shh and Nkx2.2 show partial colocalization from E9.5 to E10.5, but distinct separation at E11.5. (G-O) Combined in situ hybridization of Patched and Gli1 with immunohistochemistry for Nkx6.1 and Nkx2.2 show that Patched and Gli1 mRNA were expressed in D2 to D4 domains, especially enriched in the D3 region. Scale bars: (A) 25 μm, applied to A, D; (B) 50 μm, applied to B-C and E-F; (G) 100 μm, applied to G-O.