Fig. 4

Overexpression of HOTAIR does not induce GLI1 expression and prevents SHH-mediated induction of GLI1. Dermal fibroblasts were transduced with lentiviruses containing a scramble or HOTAIR sequence. RNA and protein were extracted from these cells. a GLI1 and NID protein levels were analysed by western blot. β-actin was used as a loading control. GLI1 (b) and PTCH1 (c) transcript levels were analysed. Graphs represent the mean and standard error for three independent experiments. RNA was extracted from scramble and HOTAIR-expressing fibroblasts. In addition, the scramble and HOTAIR-expressing fibroblasts were transfected with GLI2 siRNA. A scramble control siRNA was transfected as control. PTCH1 (d) transcript levels were assessed. Graphs represent the mean and standard error for three independent experiments. e RNA was extracted from serum depleted scramble and HOTAIR-expressing fibroblasts. In addition scramble and HOTAIR-expressing fibroblasts were stimulated with SHH ligand for 24 h. GLI1 transcript levels were analysed. Graph represents the mean and standard error for three independent experiments. f Scramble and HOTAIR-expressing fibroblasts were stained for an antibody specific for acetylated α-tubulin and visualised with an Alexa 594-conjugated secondary antibody. DAPI was used to visualise the nuclei (blue). The images are individual cells at a magnification of × 63 with a wider field of view in the insert. Graph represents mean cilium lengths from 4 independent experiments where 30 cells were measured. *p < 0.05, **p < 0.01, ***p < 0.001