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. 2008 Mar 21;105(12):4838–4843. doi: 10.1073/pnas.0712216105

Fig. 1.

Fig. 1.

Hh signaling inhibits p53 activity and down-regulates p53. (a and b) A WT (SMO-WT) or mutant (SMO-M1 or SMO-M2) Smo expression plasmid was transfected into Saos-2 cells (a) or C3H10T1/2 cells (b), with (+p53) or without the p53 expression plasmid and with the RGC-luc reporter plasmid containing synthetic p53-binding sequences (39). Luciferase activity was measured 48 h after transfection. The histogram shows the means of three independent experiments, and error bars show standard deviations (SDs). (c) p53 (+p53) and gD-tagged Smo expression plasmids (SMO-WT, SMO-M1, and SMO-M2) were transfected into C3H10T1/2 cells, and the amount of p53 was determined by immunoblotting. The Smo protein level was determined by using an anti-gD antibody. (d) The Smo expression plasmid was transfected into C3H10T1/2 cells. Forty-eight hours after transfection, p53 was detected by immunoprecipitation (IP), using anti-p53 (FL-393) antibody, followed by immunoblotting with a different anti-p53 (Pab246) antibody. (e) C3H10T1/2 cells were treated with the indicated amounts of mouse sonic hedgehog N-terminal peptide (Shh-N; R&D Systems) for 48 h. The amount of p53 was determined as described in d.