Supplementary Figure 3

(a) Confocal z sections and xy sections of ZEG;Crect limb buds at 17 som. stage (E9.0) (top) and 25 som. stage (E9.5) (bottom) visualising GFP (green) and DAPI (blue). (b) Cartilage (Alcian Blue) and bone (Alizarian Red) staining of E18.5 WT and βcat^f/f;Crect mutant forelimb (top panels) and hindlimb (bottom panels) skeletons. (c) Confocal z sections (top panels) and xy sections (bottom panels) of control βcat^f/f and βcat^f/f ;Crect mutant limb bud ectoderm visualising β-catenin (red) and DAPI (blue). (d) Confocal xy sections of control βcat^f/f and βcat^f/f;Crect mutant limb bud ectoderm expressing Tcf/Lef::H2B-Venus. Shown are GFP (green) and DAPI (blue). (e) Histogram representing percentage GFP-positive nuclei in control βcat^f/f and βcat^f/f;Crect mutant limb bud ectodermal cells expressing Tcf/Lef::H2B-Venus (p = 1.7 × 10⁻⁵ (Student’s t-test); n = 3 19–25 som. embryos). (f) Confocal projection of control βcat^f/f and βcat^f/f ;Crect mutant limb bud ectoderm visualizing phospho-histone H3 (red), Caspase3 (green), and DAPI (blue). (g) Histogram representing percentage of pHH3-positive cells in control βcat^f/f and βcat^f/f;Crect mutant limb bud ectodermal cells (p = 0.8956). (h) Histogram representing number of Caspase3-positive cells in control βcat^f/f and βcat^f/f ;Crect mutant limb bud ectodermal cells (p = 0.8413). (i) Histogram representing percentage of pHH3-positive cells in control βcat^f/f and βcat^f/f ;Crect mutant limb bud mesodermal cells (p = 0.9671). (j) Histogram representing number of Caspase3-positive cells in control βcat^f/f and βcat^f/f ;Crect mutant limb bud mesodermal cells (p = 0.7691). (For g–j n = 3 19–21 som. embryos per condition, Student’s t-test) (k) Fgf10 expression in control βcat^f/f and βcat^f/f;Crect mutant limb buds at 22 som. (E9.25). (l) Fgf10 expression in control βcat^f/f and βcat^f/f ;Crect mutant limb buds at E10.5. Scale bar indicates 10 μm (a,c,d), 100 μm (f,k,l), 1 mm (b). Error bars indicate SEM. Asterisk indicates P < 0.05.