Figure 4
From: Gene expression profiles during tissue remodeling following bladder outlet obstruction
DBP-mediated gene expression under hypoxic conditions in human bladder smooth muscle cells. (A) DBP protein levels in human bladder smooth muscle cells under hypoxic conditions. BdSMC cells were transfected with siDBP and cultured with CoCl2 solution for 24 h (0 or 500 mM). Western blotting was performed with anti-DBP and anti-HIF1α antibodies. (B) mRNA levels of DBP-knockdown BdSMC cells under hypoxic conditions. BdSMC cells were transfected with siDBP and treated with CoCl2 for 24 h. RT-qPCR was used to measure gene expression. Measurements show average values of 3 independent measurements, which were normalized to ACTB mRNA expression. **P < 0.01, *P < 0.05. (C) A schematic of gene loci of human PER1, NOS3 and AQP3. Locations of DBP-binding sequences are shown as red bars. Blue arrows indicate primers for ChIP assay. (D) ChIP assay with anti-DBP antibody using BdSMC cell lysate. BdSMC cells were cultured with 500 mM CoCl2 solution for 0 h or 4 h. Immunoprecipitated DNA fragments were quantified by qPCR. Each primer set is shown in Table S3. Data are means ± SEM for three independent experiments. *P < 0.05.
