Shh and FGF8 act on distinct aspects of mDA neuron differentiation.

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Shh and FGF8 act on distinct aspects of mDA neuron differentiation. Shh and FGF8 act on distinct aspects of mDA neuron differentiation. (A) Experimental scheme for data in B and C. (B) qPCR analysis of mouse d6 MD cultures for the ventral mesencephalic regulator genes Lmx1a, Foxa2, Msx1 and Shh and the isthmus-expressed midbrain patterning genes Wnt1, En1 and En2. (C) qPCR analysis at d9 MD for Wnt1, En1 and En2. PD0325901 (PD) was added for 2 days from d0-2, whereas Shh and/or FGF8 were added from d5. (D) qPCR analysis of Helt, Gata2 and Gad1 in d5 and d9 MD cultures. Expression levels were examined in control MD cultures with no added factors, control cultures treated with Shh between d5 and d9 alone, as well as in cultures treated with PD at d0-2 followed by Shh between d5 and d9 MD. Data in B and C are mean±s.e.m. of two experiments performed in duplicate, whereas data in D are mean± s.e.m. of triplicate cultures from one representative experiment. Ines Jaeger et al. Development 2011;138:4363-4374 © 2011.