FIGURE 9.

The recruitment of ACTN4 and GR with GR transactivated promoters in HPCs. A, diagrams of mapped GREs and PCR primer sets in GR-transactivated genes. B–E, HPCs were treated with vehicle or dexamethasone (Dex) for 4 h, followed by ChIP assays with anti-ACTN4 and anti-GR antibodies. Immunoprecipitated (IP) chromatin was analyzed by qPCR using the primers flanking the mapped putative GR-binding sites (ENCODE) on the indicated promoters. F, control and ACTN4 knockdown HPCs were treated with vehicle or dexamethasone for 8 h prior to harvest. Total RNA was prepared, and quantitative RT-PCR analysis was conducted using gene-specific primers. G–J, HPCs stably expressing shCtrl or shACTN4 were treated with vehicle or dexamethasone for 4 h prior to harvest. The relative recruitment of ACTN4 and GR to SERPINE1 (4), ANGPTL4 (4), CCL20 (4), and SAA1 (17) promoters was determined by normalizing the PCR products from shACTN4 cells to that of shCtrl cells. *, p < 0.05; **, p < 0.01; ***, p < 0.001; N.S., not significant.