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Figure 7 | Molecular Autism

Figure 7

From: Differential recruitment of coregulators to the RORA promoter adds another layer of complexity to gene (dys) regulation by sex hormones in autism

Figure 7

ChIP-reChIP analysis of RORA coregulators binding to the CYP19A1 promoter region. (A) Schematic diagram showing the upstream region of the CYP19A1 gene (edited from the UCSC Genome Browser). Potential RORA binding sites are labeled (RORAbs = RORA potential binding site). (B) Sequential chromatin immunoprecipitation (ChIP-reChIP) followed by qPCR analysis was conducted to determine whether NCOA1, NCOA5, SUMO1, or FHL2 interact with RORA in the CYP19A1 promoter region. RORAbs-II and III are located adjacent to each other and cannot be analyzed separately by qPCR analysis, and are thus shown as RORAbs-II/III. Chromatin was isolated from SH-SY5Y cells and first immunoprecipitated with anti-RORA. The RORA-immunoprecipitated chromatin was washed to remove anti-RORA, then free chromatin was re-immunoprecipitated with anti-NCOA1, anti-NCOA5, anti-SUMO1, anti-FHL2, or nonspecific IgG, and a no-second-antibody control was also included. The enrichment of RORA binding sites in the CYP19A1 promoter region in each re-ChIP sample was then determined using qPCR analysis (n = 3) and normalized with reference to the no-second-antibody control. Error bars indicate SEM. #Undetectable. ChIP-reChIP, sequential chromatin immunoprecipitation; SEM, standard error of the mean.

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