Expression analysis of single-transgenic mice for the ZFHD1::mSTAT1 or the ACT::DBD-TAD construct. Mice transgenic for the target construct ZFHD1::mSTAT1 (line #82) were crossed with non-transgenic mice. PEF were isolated from individual embryos at day 14, transfected with pEF::DBD-TAD, stimulated with 100 nM rapalog for 24 h or left untreated and subjected to RT-PCR analysis for transgenic STAT1 expression; cyclophilin was used as endogenous control (A). PEF were isolated from each embryo separately of transgenic mice (ACT::DBD-TAD, line #1 and EF::DBD-TAD, line #103), lysed and subjected to WB for analyzing expression of TAD compared to MEF line S2RS #8. Loading was controlled with p65/NFκB (B). Protein was isolated from different organs of transgenic mice and WB was performed detecting TAD subunit. As loading control a SDS-PAGE was performed and stained with Coomassie (C).