Fig. 3

Deficient sonic hedgehog (Hh) pathway in term PE hCV-MSCs. A–C hCV-MSCs were starved for 24 h, stimulated with smoothened agonist (SAG, 400 nM) for 24 h and stained for the hedgehog marker Smo (green), cilia markers Arl13b (red), and pericentrin (red). Line-scan analyses of fluorescent Smo are shown as indicated for 1st trimester (A), term control and term PE hCV-MSCs (B). A, B Each point on the graph represents the mean fluorescence intensity and the results are presented as mean ± SEM (n = 30 cilia, pooled from three independent experiments). Unpaired Mann-Whitney U test was used. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. C Representatives of treated hCV-MSCs are shown. Scale: 10 μm. D–F Relative gene expression levels of GLI1 (D), SMO (E), and PTCH1 (F). Untreated term hCV-MSCs served as reference sample and GAPDH is used as endogenous control. Student’s t test was used. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001