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. 2017 Mar 24;8:14685. doi: 10.1038/ncomms14685

Figure 1. Temporal delay of Hox protein expression in postmitotic motor neurons.

Figure 1

(a) Schematic illustration of protocol to generate spinal motor neurons from embryonic stem cells (ESCs). RA: retinoic acid. SAG: smoothened agonist. NE: neural epithelium. pMN: motor neuron progenitors. MNs: motor neurons. (b) Rapid induction of Hoxa1–Hoxa5 gene expression upon RA/SAG induction. Heat map represents the intensity of gene expression along the Hoxa cluster at five time-points during motor neuron differentiation by Quantile analysis. (c) Quantitative PCR (qPCR) analysis of Hoxa5 during MN differentiation. mRNA levels were normalized against Day4 pMN cells (mean±s.d., n=3 independent experiments). FC-fold change. (d) Immunodetection of Olig2, Hb9 and Hoxa5 in EBs under RA/SAG conditions. Scale bar represents 50 μm. (e,f) Comparisons of Olig2, Hb9 and Hoxa5 mRNA and protein expression by qPCR and immunostaining. mRNA/protein levels were normalized against Day 3 EBs (mean±s.d., n=3 independent experiments). Only Hoxa5 exhibits a significant delay of ∼72 h between gene and protein expression (f). FC-fold change, RO-ratio.