Figure 5.

Effects of an inhibitor and an angonist of hedgehog signaling. (A) Real-time RT-PCR assay for rat-specific Col2a1 mRNA. RNA was recovered from the regenerated meniscus 2 weeks after the various treatments, including injection of PBS, hMSCs, hMSCs plus 0.1% DMSO (as a control of cyclopamine), hMSCs plus cyclopamine (50 μL of a 20 μM in 0.1% DMSO), 100 nM or 500 nM Smoothened agonist (SAG), or fibroblasts. Values are mean with lower and upper limit of 95 % CI of the relative quantities (RQ) normalized to rat-specific GAPDH; n = 4 for each group. + p=0.005, ++ p= 0.002 (one-way ANOVA followed by Bonferroni post-tests. P values less than 0.0033 were considered to be statistically significant.) (B, C) Real-time RT-PCR assay for rat-specific Ptc1 (B) and Gli1 (C), downstream targets of hedgehog signaling. RNA was recovered from the regenerated meniscus 3 days after the various treatments. Values are mean with lower and upper limit of 95 % CI of the relative quantities (RQ) normalized to rat-specific GAPDH; n = 4 for each group. + p=0.004, ++ p= 0.002 in B, ++ p=0.002, +++ p=0.001 in C (one-way ANOVA followed by Bonferroni post-tests. P values less than 0.005 were considered to be statistically significant.) (D) Representative macroscopic photographs of the meniscus 2 weeks after the various treatments. Scale bar: 1 mm. (E) Area of the total meniscus 2 weeks after the various treatments. Values are mean with lower and upper limit of 95 % CI; n = 4 for each group (one-way ANOVA followed by Bonferroni post-tests. P values less than 0.0083 were considered to be statistically significant.)