Fig. 7. Decreased noradrenergic and increased cholinergic traits in Isl1 hypomorphic mutant embryos.

a, b GSEA showing downregulation of a gene set representing noradrenergic differentiation, but upregulation of a gene set representing cholinergic differentiation in Isl1 hypomorphic sympathetic ganglia. Normalized enrichment scores (NESs) and p-values are shown. c, d qPCR showing relative mRNA expression of noradrenergic and cholinergic neuron genes in Isl1 hypomorphic and control sympathetic ganglia at E14.5 and E17.5. Error bars represent the s.d., n = 3, two-tailed t-test. e–r Verification of selected genes associated with cholinergic neurons (Ret, Vip, Sst, Tlx3) and noradrenergic neurons (Hmx1, Ntrk1, TH) by in situ hybridization (e–j, m, n) and immunostaining (k, l, o–r) in E14.5 control and Isl1 hypomorphic sympathetic ganglia. Scale bar, 100 µm, ns = not significant, * p < 0.05, or ** p < 0.01