spacer gif spacer gif spacer gif spacer gif ARCHIVE ANNOUNCEMENT! spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

Right arrow Help viewing high resolution images
Right arrow Return to article
(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.


Figure 6


Fig. 6. AP1 directly binds to the regulatory regions of AGL24, SVP and SOC1. (A) Schematic of the genomic regions of Arabidopsis AGL24, SVP and SOC1. Bent arrows denote translational start sites and stop codons. Exons and introns are shown by black and white boxes, respectively. The arrowheads indicate the sites containing either single mismatch or perfect match from the consensus binding sequence (CArG box) for MADS-domain proteins. The hatched boxes represent the DNA fragments amplified in the ChIP assay. (B) Western analysis of nuclear extracts from inflorescences (i) of ap1-1, and inflorescences (i) and leaves (l) of wild-type plants probed with the purified AP1 antibody. AP1 protein was only detectable in wild-type inflorescences. (C) Western analysis of the specificity of anti-AP1 serum in the ChIP procedure. After sonication, the supernatant containing solubilized chromatin from wild-type inflorescence served as an input for immunoprecipitation either with IgG(-) or with anti-AP1 serum (+). Anti-AP1 serum could specifically precipitate AP1 protein. (D) Western analysis of the specificity of anti-AP1 serum to precipitate AP1-GR fusion protein in the ChIP procedure. After sonication, the supernatant containing solubilized chromatin from inflorescences of wild-type and ap1-1 35S:AP1-GR (Dex- or Mock-treated) plants served as an input for immunoprecipitation either with IgG(-) or with anti-AP1 serum (+). Anti-AP1 serum could specifically precipitate AP1-GR protein. (E-G) ChIP analysis of AP1 binding to regulatory sequences of AGL24 (E), SVP (F) and SOC1 (G). Real-time PCR assay of immunoprecipitated DNAs was conducted in triplicate. Relative enrichment of each target DNA fragment was calculated first by normalizing the amount of a target DNA fragment against a TUB2 genomic fragment, and then by normalizing the value for anti-AP1 serum against the value for IgG. The enrichment of an ACTIN 2/7 gene fragment was used as a negative control. Error bars indicate the standard error of the mean.





Right arrow Return to article