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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. 2.

Fig. 2

Conditioned media from cells transfected with full length HH demonstrates isoform-specific autoprocessing and secretion. (A) Light2 cells were assayed for luciferase activity after 48 h treatment with conditioned media (1:4 in medium with 0.5% FBS, with and without cyclopamine as indicated) generated from 293T cells transfected with full length HH cDNA and cultured for 48 h in medium with 2% FBS. EcR Shh was used as a positive control for secreted HH. (B) Gli1 protein levels were determine by Western blot on 3T3 cells after 48 h treatment with conditioned media (1:4 in medium with 0.5% FBS). Quantified ratio of Gli1/GAPDH protein is shown below GAPDH for each lane. (C) HH production and secretion was determined by Western blot of cell lysates (CL) and conditioned media (CM) from 293T transfected cells for HH and cultured in medium with 2% FBS for 48 h, probed with 5E1 anti-HH or anti-DHH primary antibody as indicated. TCA denotes TCA precipitated CM.