Project description:This experiment describes gene expression after the activation of APETALA1-GR, to study and identify AP1 target genes. We used a pAP1:AP1-GR ap1 cal line to induce a synchronized response activating the AP1-GR fusion protein in ap1 cal inflorescence-like meristems through dexamethasone treatment. Tissue samples were collected immediately after the treatment, as well as subsequent timepoints. The expression profiles of the individual samples were then analyzed by gene expression profiling using whole-genome oligonucleotide arrays (Agilent, custom-commercial)
Project description:This experiment describes gene expression after the activation of APETALA1-GR, to study and identify AP1 target genes. We used a pAP1:AP1-GR ap1 cal line to induce a synchronized response activating the AP1-GR fusion protein in ap1 cal inflorescence-like meristems through dexamethasone treatment. Tissue samples were collected immediately after the treatment, as well as subsequent timepoints. The expression profiles of the individual samples were then analyzed by gene expression profiling using whole-genome oligonucleotide arrays (Agilent, custom-commercial) Four sets of biologically independent tissue samples were collect before DEX-induction (day 0, 0d), as well as at 2 days, 4 days and 8 days after the treatment. Samples for each timepoint and biological replicate were co-hybridized as indicated; one sample was labeled with Cy3 and the other with Cy5. The dyes used for labeling RNA from a given timepoint were switched for two of the replicate experiments, to reduce dye-related artifacts. This experimental setup resulted in a total of 3 hybridizations per set (0days vs 2days, 2days vs 4days, and 4days vs 8days), and an even number (2) of hybridizations of each dye polarity per comparison.
Project description:This experiment describes gene expression after the activation of APETALA1-GR, to study and identify AP1 target genes. We used a 35S:AP1-GR ap1 cal line to induce a synchronized response activating the AP1-GR fusion protein in ap1 cal inflorescence-like meristems through dexamethasone or dexamethasone+cycloheximide treatment. Tissue samples were collected at 3hrs after the treatment. The expression profiles of the individual samples were then analyzed by gene expression profiling using whole-genome oligonucleotide arrays (Agilent, custom-commercial). We treated inflorescences of 35S:AP1-GR ap1-1 cal-1 plants with a dexamethasone-containing or a mock solution, or with identical solutions that contained in addition 10 M-NM-<M cycloheximide. Tissue was collected 3 hours after the treatment. Samples from each of the four biological replicates resulted in a set of four hybridization pairs: Mock vs. Dex, Mock vs. Chx, Mock vs. Dex+Chx, and Chx vs. Dex+Chx. Dye polarities were switched between biological replicates.
Project description:Plant inflorescence-to-floral phase transition is an important developmental stage, in which floral cell identities and many traits of reproductive organs are determined. Two MADS-domain transcription factors, APETALA1 (AP1) and CAULIFLOWER (CAL), have been known as master regulators controlling the early stage of the phase transition in Arabidopsis. In plants with loss-of-function alleles of ap1 and cal double mutations, flower development is heavily delayed at the flower initiation stage and accumulate a large number of inflorescence-like meristem cells compared to wild-type plants, resulting in a cauliflower-like phenotype. To facilitate investigation on molecular mechanisms during inflorescence-to-floral phase transition, an inducible system of synchronized floral development has been developed, in which ap1,cal inflorescence-like meristem cells express a fusion protein of AP1 and the hormone-binding domain of the rat glucocorticoid receptor (GR) driven by 35S constitutive promoter. When inflorescences of 35S:AP1-GR ap1,cal plants are treated by steroid hormone dexamethasone as the activator to allow the AP1-GR fusion protein translocate into nucleus, inflorescence-to-floral phase transition is triggered and plants start to produce hundreds of relatively synchronized floral buds. To explore molecular basis at early stage of flower development in Arabidopsis, we used the inducible system of synchronized floral development (35S:AP1-GR ap1,cal) to profile transcriptome change of meristem cells during inflorescence-to-floral phase transition by strand-specific RNA-sequencing.
Project description:This experiment describes gene expression during early Arabidopsis flower development. We used a 35S:AP1-GR ap1 cal line to induce synchronized flower development by specifically activating the AP1-GR fusion protein in ap1 cal inflorescence-like meristems through dexamethasone treatment. Tissue samples were collected immediately after the treatment, as well as at one-day intervals for the following five days. The expression profiles of the individual samples were then analyzed by gene expression profiling using whole-genome microarrays (Operon). Keywords: time course