TGF -β enhances GLI2 and GLI1 expression in human adrenocortical carcinoma cells. (A) NCI-H295R cells were treated with 10 ng/ml TGF-β for 4, 8, and 24 h and GLI2 and GLI1 expression was analyzed by quantitative RT-PCR using 18S rRNA as an internal control. Data are presented as mean ± SD, n = 4–6 replicates, pooled from two experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001. (B) Western blots of lysates from NCI-H295R cells probed for GLI1 and GLI2 (and ACTIN as a loading control), after treatment with 10 ng/ml TGF-β and 50 µg/ml cycloheximide (CHX) for 48 h. The arrows indicate the bands of GLI2 and GLI1.