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. 2015 Jan 13;5:7750. doi: 10.1038/srep07750

Figure 1. egr4 is a downstream target of Pax3 and Zic1.

Figure 1

(a) Whole-mount in situ hybridization for egr4, pax3 and zic1. The DIG-labeled probes were used alone or in combination (pax3/egr4 and zic1/egr4). Dorsal views, anterior to top. Scale bar, 300 μm. (b) Injection in one blastomere at the 2-cell stage of MOs to block Pax3 (pax3MO; 40 ng) or Zic1 (zic1MO; 40 ng) function resulted in a reduction of egr4 expression. The injected side is to the right as indicated by the presence of the lineage tracer (Red-Gal). Dorsal views, anterior to top. Scale bar, 300 μm. (c) The graph indicates the percentage of embryos with normal (white) or reduced/lost (red) egr4 expression. The number of embryos analyzed is indicated on top of each bar. (d) mRNA encoding pax3GR and zic1GR (100 pg each), alone or in combination were injected into both blastomeres in the animal pole at the 2-cell stage. At the blastula stage (stage 9), animal cap explants were dissected and cultured for 8 hours in the presence of dexamethasone (DEX). In some samples, cyclohexamide (CHX) was used to block protein synthesis. (e) egr4 expression in pax3GR and zic1GR injected animal cap explants analyzed by qPCR.