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. 2013 May 1;8(5):e63143. doi: 10.1371/journal.pone.0063143

Figure 1. Itm2a is a novel Pax3 target.

Figure 1

A–C, Whole mount in situ hybridization with an Itm2a antisense riboprobe at E9.5 (A), E10.5 (B) and E11.75 (C). D–F’, Whole mount in situ hybridization with an Itm2a antisense riboprobe on wild type (WT) (D, D’), Pax3nLacZ/nLacZ (E, E’) and Pax3Pax3-En/+ (F, F’) embryos at E10.5. D’–F’ show close-ups of the interlimb somite region. As expected, the modification of Itma transcription in the mutant is restricted to Pax3-expressing cells. G–H, Whole mount in situ hybridization with an Itm2a antisense riboprobe on wild type (WT) and Pax3PAX3-FKHR/+ embryos at E11.5. I, Real-time quantitative PCR using primers for the putative Pax3 binding site identified in [19] in the Itm2a sequence at +3.6 kb. The 5 Pax3-binding sites in the P34 transgene [21] and a functional Pax3 site at −57.5 kb from the Myf5 gene [11] provide positive controls. A Myf5 flanking sequence at −256 kb that does not bind Pax3 [11] provides a negative control. Results are expressed as a percentage of PCR signal on input DNA, showing enrichment after Pax3 immunoprecipitation, or with an IgG antibody.