Loss of mFu does not impact Hh-dependent patterning of the ventral neural tube. (A) Specification of neural tube markers is shown for Shh (a and b), Nkx2.2 (c and d), MNR2 (e and f), Isl1 (g and h), Lim3 (i and j), Lim1/2 (k and l), Pax6 (m and n), and Pax7 (o and p). Frozen sections of E10.5 embryos were stained with monoclonal antibodies against the indicated antigens and counterstained with DAPI, and photos were taken at amagnification of ×20. No defects were observed in the specification of any neuronal population in fu−/− mice. (B) Specification of serotonergic neurons is normal in fu−/− embryos. Embryos were collected at E13.5 and stained for serotonergic specified neurons with antibodies against 5-HT. Whole-mount immunostaining of the ventral hindbrain is shown, with the rostral-most region at the top. Specification of serotonergic neurons is seen in two clusters consisting of rhombomeres 2 and 3 and 5 to 7, whereas rhombomere 4 is characteristically negative for 5-HT.