Fig. 7.

Chromatin immunoprecipitation (ChIP) assay indicates that IL-6 induced phospho-Stat3 is recruited to a putative Stat3 site in the rat apelin promoter in primary cultured rat ileal cells. Top: schematic of the proximal promoter region depicting location of a putative Stat3 binding site and start sites of distal and proximal primers (arrows) used in PCR amplification. −289 and −88 bp = start sites. ChIP assays were done using chromatin isolated from vehicle (lane 1) or IL-6-treated (lane 3) primary cultured rat ileal cells. Chromatin was cross-linked, sheared, and immunoprecipitated by a phospho-Stat3 antibody. Immunoprecipitations were followed by PCR amplification of the promoter region containing the putative Stat3 binding site (−198/−195 bp). Positive input DNA (lane 2) used for PCR was 2% of the amount used for immunoprecipitation. DNA size markers are shown.