Fig. 2

The Hedgehog pathway could be restored at protein and RNA level in primary cilia of obese ASCs by pretreatment with MLN or PD. a The working schedule. b, d Cells were stained against Smo (green), pericentrin (red), Arl13b (red), and DAPI (blue). Representatives are shown. Scale bar, 10 μm. Insets depict shifted overlays. Scale bar, 2.5 μm. c, e ob-ASCvis (C) and ob-ASCsub (E) were treated as indicated and stained for the evaluation of the cilium length. The results are from three experiments (n = 73–80 cilia for each condition in each group) and presented as median ± min/max whiskers in box plots. f Line-scan analyses of fluorescent Smo are shown for ob-ASCvis (left) and ob-ASCsub (right) treated with SAG and indicated inhibitors for 24 h. Each point on the graph represents the mean fluorescence intensity (mean ± SEM) based on three experiments (n = 30 cilia). g The gene levels of GLI1, PTCH1, SMO, and NANOG are shown for visceral obese ASCs treated as indicated in a. The results are from three experiments, presented as mean ± SEM. Unpaired Mann-Whitney U test for c, e, and f. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Student’s t test for g