DEV02298 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Differences in fore- and hindlimb outgrowth visualized by skeletal analysis of wild-type and β-cat^ΔPrx1/– limbs. (A-B′) Alcian Blue-stained skeletal elements of wild-type (A) and β-cat^ΔPrx1/– (A′) forelimbs, and wild-type (B) and β-cat^ΔPrx1/– (B′) hindlimbs at E13.5 (n=5). (C-D′) Skeletal elements of wild-type (C) and β-cat^ΔPrx1/– (C′) forelimbs, and wild-type (D) and β-cat^ΔPrx1/– (D′) hindlimbs at E14.5 (n=4). Images are taken at the same magnification. Note that hindlimbs are more developed than forelimbs.

• Supplemental Figure 2 -

  Fig. S2. Analysis of Prx1Cre activity using ZAP reporter mice. (A-C′) Alkaline phosphatase staining on Prx1Cre;ZAP embryos at somite stage 12 (A), 15 (B,B′), and 18 (C,C′). Close up of forelimb region at somite stage 15 (B′) and 18 (C′).

• Supplemental Figure 3 -

  Fig. S3. Altered expression of ectodermal and mesenchymal markers in β-cat^Δex3Prx1/+ and β-cat^ΔPrx1/– limbs. (A-E′′) Section in situ hybridisation showing Tbx5, Fgf8, Twist, Tcf1 and Lef1 expression in wild-type (A-E), β-cat^Δex3Prx1/+ (A′-E′), and β-cat^ΔPrx1/– (A′′-E′′) forelimbs at E11.5. Note that β-cat^Δex3Prx1/+ limbs are expanded, whereas β-cat^ΔPrx1/– limbs are reduced in size along the DV axis, in comparison with wild-type limbs.

• Supplemental Figure 4 -

  Fig. S4. In vitro evaluation of potential β-catenin targets. Semi-quantitative RT-PCR (5× dilutions), from AdCre- and AdGfp-infected β-cat ex3fl/ex3fl micromass cultures (12 hours after infection), for Bmp4, Bmp7, Emx2, Lef1, Tcf1, Bmp2, Lmx1b, Twist1, Fgf10, Tbx5 and Sox9, in comparison with Gapdh levels. Cycle number is indicated on the right.