Figure 4.

Analysis of our Tet-On TA in embryos and larvae. (A) Mothers carrying two copies of both the actin5C-rtTA-M2-alt transgene and the TetO-luciferase transgene were fed yeast paste supplemented with dox (100 μg/ml) for 48 h (+), or dox-free yeast paste (−). Embryos of 6–18 h were dechorinated, fixed, and incubated with an antiluciferase mAb overnight at 4°C. Antibody complexes were detected with an horseradish peroxidase-conjugated secondary antibody and Ni- diaminobenzidine staining. (B–D) Wandering third-instar larvae carrying one copy of both the actin5C-rtTA-M2-alt and the TetO-LacZ transgenes were dissected, fixed, and stained with X-gal. Brains (B), wing discs (C), and leg discs (D) from larvae fed dox-containing (+) food (10 μg/ml) or regular food (−) were compared. No staining was detected in wild-type larvae or larvae carrying only the TetO-LacZ transgene (data not shown).