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. Author manuscript; available in PMC: 2008 Dec 8.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2007 May 3;27(8):1760–1767. doi: 10.1161/ATVBAHA.107.144881

Figure 3.

Figure 3

ChIP assay for the NonO or hnRNP-K proteins and P4Hα1 promoter interaction. HASMC nuclei were extracted by using the Nuclear and Cytoplasmic Extraction Reagents kit (Pierce). HASMCs were treated with or without 100 ng/mL TNF-α for 8 hours before nuclear extraction. The ChIP assay was then performed. Anti-human NonO or hnRNP-K antibodies were used in immunoprecipitation. The recovered protein-bound DNA was amplified by PCR with primers that cover the −60 to +104bp region of the P4Hα1 promoter. The PCR products were subjected to electrophoresis in 1.5% agarose gel. NonO-P4Hα1 promoter binding was only found in the TNF-α–treated cells. However, hnRNP-K did not appear to directly bind with the P4Hα1 promoter in the ChIP assay.