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. 2014 Apr;1843(4):703–714. doi: 10.1016/j.bbamcr.2014.01.003

Fig. 5.

Fig. 5

ULK3 participates in regulation of GLI1/2 transcriptional activators in ASCs. (A) Expression of genes encoding mediators of the Shh signaling pathway was analyzed using RT-PCR in ASCs isolated from four donors. (B) Primary cilia (red) and nuclei (blue) were stained using antibody against acetylated tubulin and DAPI, respectively. (C) Levels of GLI1 (a), PTCH1 (b), GLI2 (c) and ULK3 (d) mRNAs were measured by qPCR in ASCs stimulated with Shh or TGF-β1. (D) ASCs were transfected with Neg. or ULK3-specific siRNAs. The levels of ULK3 (a), GLI2 (b) and GLI1 (c) transcripts were measured by qPCR; (d) GLI1 and GLI2 proteins were analyzed by immuno-blotting. (E) ASCs were transfected with Neg. or ULK3-specific siRNAs and 24 h later treated with TGF-β1 for 24 h. The levels of ULK3 (a), GLI2 (b) and GLI1 (c) transcripts were measured by qPCR. (F) (a) GLI2 and GLI1 proteins were analyzed by WB in cells transfected with Neg. or ULK3-specific siRNAs and induced with TGF-β1 for 30 h. GLI2, GLI1 and GAPDH images were quantified. Normalized GLI2 (b) and GLI1 (c) protein levels were set as 1 in cells transfected with Neg. siRNA. Data of qPCR and quantified WBs are presented as an average mean ± S.D.; *—p < 0.05, **—p < 0.01, ***—p < 0.001, n = 3.