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. Author manuscript; available in PMC: 2015 Feb 1.
Published in final edited form as: Mech Dev. 2013 Dec 14;131:137–149. doi: 10.1016/j.mod.2013.12.002

Fig. 1.

Fig. 1

Direct transfection of FL-HH cDNA generates a Gli1 response. (A) 3T3 cells were transfected with full-length HH constructs and after 48 h culture in 0.5% FBS medium, mRNA for HH-responsive genes Gli1 and Ptch1 was quantified by qPCR. mRNA expression levels are shown as a % of positive control (N-Shh) expression. Protein levels of Gli1 were confirmed by Western blot (B) of lysates from transfected 3T3 cells. Quantified ratio of Gli1/GAPDH protein is shown below GAPDH for each lane. (C) Light2 cells transfected with full length HH constructs were assayed for luciferase activity after 48 h culture in 0.5% FBS medium. Luciferase activity is shown normalised to 100% of positive control (N-Shh). (D) CH310T1/2 cells and TM3 cells (E) were transfected with full length HH constructs and Gli1 and Ptch1 mRNA assayed by qPCR after 48 h culture in 0.5% FBS medium. (F) Gli1 protein induction in transfected TM3 cells was confirmed by Western blot on cell lysates following HH transfection and 48 h culture in 0.5% FBS medium.