Figure 5

Presence of C/EBPβ and SUMO-1 on the NOS2 promoter. Primary astrocytes were treated with nothing (C), LPS (L), or LPS+NA (LN) for 4 hours, then ChIP analysis conducted. (A) Two fragments of the rat NOS2 promoter containing the proximal C/EBP-2 (top) or C/EBP-3 (bottom) binding sites were amplified by PCR after immunoprecipitation using (B) anti-C/EBPβ antibody or (C) anti-SUMO-1 antibody alone or followed by anti-C/EBPβ antibody. The products were separated through 2% agarose gels and their accuracy was verified by comparison to input DNA ("in", 5% of total input). The images shown are inverted versions (Adobe Photoshop) of the original gel images and are representative of results obtained in 2 separate experiments. (D) Lysates of astrocytes treated as indicated above were immunoprecipitated (IP) with anti-C/EBPβ antibody and Western blot (W) of the samples was probed with anti-SUMO-1 antibody.