Figure 4

AG and SEP3 expression analysis and chromatin immunoprecipitation (ChIP). Confocal Scanning Laser Microscopical (CSLM) imaging of (A) gAG:GFP (pARC422) and (B) gSEP3:GFP (pARC423) in the inflorescence. Top view (A, B) of an inflorescence with different floral bud stages (indicated by numbers). The GFP expression (green signal) is predominantly localized in the nuclei of floral meristem cells of flower buds from stage 3 onwards (comprising whorl 3 and 4 for AG, and whorl 2, 3, and 4 for SEP3, respectively). Autofluorescence is visible as red signal. (C) Anti-GFP Western blot with material from Arabidopsis WT and gAG:GFP (pARC422) plants. Protein product is detectable in transgenic plants only. Bottom panel shows the Coomassie stained gel serving as loading control. (D) Enrichment of AG target DNA after ChAP with GFP antibody and compared with pre-immune. Quantification of target DNA was done by Real-time PCR using primers corresponding to sequences in the second intron of AG. FM, floral meristem, S, sepal, IM, inflorescence meristem, WT, wild-type.