Liver specific expression of JDP2 in JDP2-transgenic and control mice. A. HA-JDP2 expressing mice responder line was generated in the pBIG bi-cistronic expression plasmid under the regulation of tetracycline response elements. The promoter co-regulates the expression of the β-galactosidase reporter together with HA-JDP2 protein. tTA-responder mice express the tetracycline activator (tTA) under the control of Liver Activating Protein promoter (LAP). tTA is expressed specifically in hepatocytes. The association of the tTA with the DNA is prevented in the presence of doxycycline thus results in shut-off transgene expression. B. Mouse tail genotyping by PCR was performed with specific oligonucleotides corresponding to the tTA driver (tTA, right panel) and JDP2-transgene (JDP2, left panel). DNA derived from either double transgenic mouse, JDP2-transgenic (lanes 1) or control wild type mouse (lanes 2) was used. Lanes 3 no DNA was added to the PCR mix. C. Western blot analysis with nuclear extract derived from the indicated tissues of JDP2-transgenic mouse (tg) and liver lysate from wild type mouse (wt). Membranes were probed with anti-JDP2 antibody (top panel) anti-HA antibody (middle panel) and anti-α-tubulin antibody (bottom panel) was used as loading control. The migration of HA-JDP2 transgene and endogenous JDP2 protein is indicated by an arrow at the right side of the top panel. D. β-galactosidase activity with cell lysate derived from the indicated tissues of JDP2-transgenic mouse. The O.D. value at 420 nm is shown. E. Western blot analysis with liver nuclear cell lysate derived from either wild type (wt) or JDP2-transgenic (tg) mice either untreated (-dox) or treated with doxycycline (+dox, 0.2 mg/ml) for 3 days. Membrane was probed sequentially with anti-JDP2 (top panel) followed by anti-α-tubulin (bottom panel) as loading control.