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Determination of BMP-2-induced restriction enzyme accessibility in the proximal Vanoxerine dihydrochloride of the Sox9 gene
Fig. 2.
Alteration in the restriction enzyme accessibility at Sox9 proximal promoter induced by BMP-2 treatment. (A) Schematic representation of the probe used to map the restriction enzyme hypersensitive sites within the Sox9 promoter by the indirect end-labeling technique. (B) Primary MEFs were treated with or without 200 ng/ml BMP-2 for 1 d. Nuclei were then purified and digested with 25–200 U of restriction enzyme/ml. Purified DNA was digested to completion with DraI. Products were detected by Southern blotting. (C) Primary MEFs were transfected with NF-YA-targeted siRNA, NF-YB-targeted siRNA, or a primary growth control siRNA for 24 h. The whole lysates were detected by immunoblotting with antibodies. (D) Primary MEFs were transfected with siRNAs or infected with adenovirus for 24 h, or pretreated with SB203580 for 30 min, followed by BMP-2 treatment for 1 d. Nuclease hypersensitivity assays were performed using NheI.