Fig 7.

Jak2 regulates the transcriptional activity of the CryAB promoter in cardiomyocytes. (A) HL-1 cells were co-transfected with CryAB-Luc plasmid along with parental vector (pcDNAV5) or pTel-Jak2. Cells were serum-starved and treated with vehicle (DMSO) or 50 μM AG490 for 24 hrs. Luciferase activities in cells treated with AG490 and co-transfected with Tel-Jak are expressed relative to the mean value of the co-transfected cells exposed to the vehicle. Renilla luciferase expression was used for normalization. Experiments have been repeated at least three times. Data are mean ± SE. *P < 0.05 and **P < 0.01. (B) Serum-starved HL-1 cells were treated for 5, 10 and 30 min. with either En-1 (50 nM) or vehicle (water) and subjected to ChIP assays. Soluble chromatin from cross-linked cells was immunoprecipitated with indicated antibodies. A non-specific IgG was used as a negative control for immunoprecipitation and the antibody alone (Ab), without lysate, served as a control for cross-contamination. Input DNA represents 10% of total chromatin used in each reaction. DNA was amplified performed with primers flanking the αBE4 sequence. Primers specific to GAPDH were used before (input) and after immunoprecipitation as a control to monitor immunoprecipitation specificity. Data shown are one representative of three separate experiments.