Figure 4: Foxc1 controls the expression of Ihh target genes by recruiting Gli2.

(a,b) Synergistic effect of Foxc1 and Ihh on Gli1 (a) and Ptch1 (b) mRNA expression in primary chondrocytes infected with adenoviruses as indicated. Data are shown as the mean±s.d. (n=3). **P<0.01 (versus Control, Foxc1 or Ihh); one-way analysis of variance (ANOVA) followed by the Tukey–Kramer test. (c,d) Synergistic effect of Foxc1 and Gli2 on Gli1 (c) and Ptch1 (d) mRNA expression in primary chondrocytes infected with adenoviruses as indicated. Data are shown as the mean±s.d. (n=3). **P<0.01 (versus Control, Foxc1 or Gli2); one-way ANOVA followed by the Tukey–Kramer test. (e,f) Total RNA was isolated from the microdissected hindlimbs of E15.5 WT and Foxc1^ch/ch littermate embryos, and Gli1 (e) and Ptch1 (f) mRNA expression was determined by RT–qPCR. Data are shown as fold expression normalized to WT (mean±s.d., n=3). **P<0.01 (versus WT); Student’s t-test. (g–i) Primary chondrocytes were infected with control, Foxc1, Gli2 or both Foxc1 and Gli2 adenoviruses and cultured for 3 days. ChIP assays were conducted using an anti-Gli2 antibody and DNA binding to gene promoters was determined by qPCR using primer pairs specific for PTHrP (g), Gli1 (h) and Ptch1 (i) gene promoters, which contain the Gli-binding element. Data are shown as the mean±s.d. (n=3). **P<0.01 (versus control, Foxc1, or Gli2); one-way ANOVA followed by the Tukey–Kramer test.