Fig. 1

TC17 validation in vitro and in vivo strategy. A 293 T cells were transfected with three plasmids containing CRE, LoxP/TetOn, and Cyp17/Flag. After exposure for 24 h with or without 1 ug/ml Dox, cell lysates were subjected to immunoblotting analysis with anti-Flag and anti-Cyp17 antibodies. B Homozygous transactivator mice were bred with homozygous responder mice. Double transgenic mice were then bred with Cyp17iCre transgenic mice [60] to produceR26-STOP-rtTA-IRES-EGFP/TRECyp17/Cyp17iCre triple transgenic mice. R26, a ubiquitous and endogenous ROSA26 promoter; rtTA, reverse Tet-controlled transcriptional activator; IRES, internal ribosome entry site; EGFP, enhanced green fluorescent protein; Dox, doxycycline