Figure 1: Rescue of Tbx4 expression in Pitx1−/− hindlimb buds.
From: Divergent transcriptional activities determine limb identity
(a) In situ hybridization showing Tbx4 mRNA expression in wild-type (Pitx1+/+) and Pitx1−/− e10.5 HL buds. Bar represents 500 μm. (b) Immunohistochemical detection of nuclear Tbx4 protein in transverse sections of e11.5 embryos of wild-type and Pitx1−/− HL buds and flank mesenchyme (fm). Bar represents 100 μm. (c) Western blot analysis of Tbx4 and α-tubulin levels in Pitx1+/+, Pitx1+/− and Pitx1−/− e11.5 HL buds. Normalized Tbx4 levels are shown relative to α-tubulin levels (n=3). (d) Schematic representation of transgenes used in this study. Transgenic activity of the Prx1 promoter was described previously37. (e) Whole-mount in situ hybridization analysis of transgene expression in e11.5 embryos. Bar represents 500 μm. (f) Quantitation of Tbx4, Tbx5 or Tbx4Q538R expression levels by reverse transcriptase–quantitative PCR performed on RNA extracted from dissected e11.5 HL buds expressed relative to Gapdh mRNA. Error bars indicate s.e.m. (n=3).
