Figure 5

The Hdac5 gene is positively regulated by Pax3 and negatively affects Foxc2. (A) Whole mount in situ hybridisation at E10.5 with an Hdac5 probe. Arrows point to expression at sites of myogenesis in the somites and forelimb bud. (B) GFP positive cells were isolated by flow cytometry (see Figure 2) from interlimb somites of Pax3^GFP/+ and Pax3^GFP/nlacZ embryos and the same numbers of cells were used for RNA preparation and quantitative PCR analysis of Hdac5 and Gapdh transcripts; mRNA levels are shown relative to Gapdh transcripts with the heterozygote control (Pax3^GFP/+) level taken as 1. Histograms represent mean fold change for two different biological samples analyzed in duplicate at each stage; error bars, standard error to the mean (SEM). (C-D) Whole mount in situ hybridisation with a Foxc2 probe on Hdac5^+/- (C) and Hdac5^-/- (D) embryos at E9.5, showing up-regulation of Foxc2 in somites in the absence of Hdac5.