The miR-23a/24-2 and miR-210 promoters are regulated by RARA. (A) RT-qPCRs directed against the miR-23a, miR-210, and miR-223 were performed in 293T cells treated with ATRA (10 nM). Results are indicated as fold change, 1 being the value obtained in the absence of treatment. (B) ChIPs performed in 293T cells using the indicated antibodies. The enriched genomic fragments were amplified using specific primers. The RARB promoter was used as a positive control. 223-A and 223-B as in Figure 2A. (C) qPCR analyses performed on DNA immunoprecipitated in panel B. (D) LUC assays performed in 293T cells transfected with promoterless pGL3b plasmid (black histograms) or the firefly luciferase reporter driven by the miR-23a/24-2 promoter (gray histograms). Cells were treated with increased doses of ATRA for 16 hours. Results are expressed as RLUs, 1 representing the value obtained with the pGL3b vector in absence of treatment. (E) LUC assays performed in 293T cells transfected with the firefly luciferase reporter driven by the miR-23a/24-2 promoter or the miR-23a/24-2 promoter mutated in the predicted PML-RARA response element. Cells were treated (gray histograms) or not (black histograms) with ATRA (10 nM) for 16 hours. Results are expressed as RLU, 1 representing the value obtained with the pGL3b vector in absence of treatment.