Fig. 3. Delayed differentiation of sympatho-adrenal precursor cells in Insm1 mutant mice. The differentiation of the sympatho-adrenal precursor cells in Insm1^lacZ/+ and Insm1^lacZ/Insm1^lacZ mice was assessed at the indicated stages using immunohistochemical analysis or in situ hybridization. (A-P) Analysis of E10.5 embryos. (A-D) Immunohistochemistry using antibodies against β-galactosidase (red), (A,B) Phox2b (green) and (C,D) Phox2a (green). (E-J) In situ hybridization using probes specific for (E,F) Hand2, (G,H) Gata3 and (I,J) Ret. (K,L) Immunohistochemistry using antibodies against β-galactosidase (red) and Th (green). (M,N) In situ hybridization using a Dbh-specific probe. (O,P) Immunohistochemistry using antibodies against β-galactosidase (red) and Tuj1 (green). There is a pronounced reduction in the number of sympatho-adrenal precursor cells that express Phox2a, Hand2, Gata3, Ret, Th or Dbh at E10.5 in Insm1^lacZ/Insm1^lacZ mice. (Q,R) Analysis of E12.5 embryos by immunohistochemistry using antibodies against β-galactosidase (red) and Th (green). (S) Quantification of β-galactosidase+ sympatho-adrenal precursor cells that co-express Th in Insm1^lacZ/+ and Insm1^lacZ/Insm1^lacZ mice at various developmental stages. (T,U) Analysis of E12.5 embryos by immunohistochemistry using antibodies against β-galactosidase (red) and Phox2a (green). (V) Quantification of the β-galactosidase+ cells in sympathetic ganglia that co-express Phox2a at various developmental stages in Insm1^lacZ/+ and Insm1^lacZ/Insm1^lacZ mice. (W,X) Analysis of E12.5 embryos by immunohistochemistry using antibodies against β-galactosidase (red) and Tuj1 (green). (Y) Quantification of β-galactosidase+ sympatho-adrenal precursor cells that co-express Tuj1 in Insm1^lacZ/+ and Insm1^lacZ/Insm1^lacZ mice at various developmental stages. Single or double asterisks in S,V,Y indicate P<0.05 and P<0.01, respectively. Scale bars: 50 µm in A; 100 µm in C,Q.