Figure 2

Integration site(s) and transgene expression analysis of rtTA-M2 transgenic rat founders. A. Southern blot analysis. Genomic DNA from four transgenic founder animals (Tg124, Tg129, Tg135, Tg144) was digested with EcoRI and hybridized with rtTA-M2 cDNA. B. Expression and function analysis of the rtTA-M2 transgene. Tail fibroblasts from the four rtTA-M2 transgenic founders were transduced with Lenti-TetO₇/CMV-EGFP and cultured in the presence of Dox (500 ng/ml). EGFP expression was observed by immunocytochemistry 48 hours post-transduction using an epifluorescent microscope and a FITC filter cube. EGFP expression was observed in transgenic founders, Tg129 and Tg144, indicating expression of the rtTA-M2 transgene and transactivation of the TetO₇/CMV-EGFP transgene. EGFP expression was not detectable in tail fibroblasts from transgenic founders Tg124 and Tg135. EGFP expression was not observed in fibroblasts from any of the transgenic founders if Dox was not added. Bar = 20 μm.