Project description:Rice plants cultivated on mountain areas are frequently exposed to acid fog and natural fog events. In this report, we analyzed the expression profiles of the rice plant with acid fog (SiAF) or neutral fog (SiNF) treatment for 1 and 7 days. Microarray results suggested that ROS generation was induced by not only SiAF, but also SiNF treatment, and it occurred in apoplast, not in organelles. Genes for defense- and stress-responses was also induced by both SiAF and SiNF treatments. The induction occurred in plants treated with SiAF for both 1 day and 7 days, and it was also detected in plants treated with SiAF for 7 days. These results suggest that both SiAF and SiNF treatments are abiotic stresses accompanying ROS generation in apoplast. Comparison between continuous fog treated shoots and control shoots. Biological replicates: 3 simulated acid fog (pH 3.0) treated (SiAF) for 1 day; 3 simulated acid fog (pH 3.0) treated (SiAF) for 7 days; 3 simulated neutral fog (SiNF) treated for 1 day; 3 simulated neutral fog (SiNF) treated for 7 days; 3 controls for 1 day; 3 controls for 7 days. Independently grown and harvested. Cultivar: Hinohikari 1 sample derived from 5 plants grown under the same conditons. Total: 18 samples, 12 comparisons.
Project description:FOG-1/CPEB and FOG-3/Tob are the terminal regulators of the sex determination in C. elegans germ cells. CPEB and Tob proteins are both translational regulators. To investigate how FOG-1 and FOG-3 regulate germ cell sex determination we sought to identify the target mRNAs. We used transgenic epitope tagged animals (3xMyc::FOG-1 and FOG-3::3xFLAG). To identify the mRNA targets of FOG-1/CPEB and FOG-3/Tob on a genome wide scale we used RNA immunoprecipitation followed by microarray analysis. We found 81 putative mRNA targets of FOG-1 and 722 putative targets of FOG-3. 76 target mRNAs were common to both FOG-1 and FOG-3.
Project description:FOG-1/CPEB and FOG-3/Tob are the terminal regulators of the sex determination in C. elegans germ cells. CPEB and Tob proteins are both translational regulators. To investigate how FOG-1 and FOG-3 regulate germ cell sex determination we sought to identify the target mRNAs. We used transgenic epitope tagged animals (3xMyc::FOG-1 and FOG-3::3xFLAG). To identify the mRNA targets of FOG-1/CPEB and FOG-3/Tob on a genome wide scale we used RNA immunoprecipitation followed by microarray analysis. We found 81 putative mRNA targets of FOG-1 and 722 putative targets of FOG-3. 76 target mRNAs were common to both FOG-1 and FOG-3. FOG-1 RNA-coimmunoprecipitation (RIP) samples were prepared as follows. Parallel RIPs were performed with wild type animals as the control IP. Worm extracts were made from wildtype and 3xMyc::FOG-1 late L3/early L4 animals. Myc affinity gel was used to immunoprecipitate 3xMyc::FOG-1 from the extracts. RNA was extracted from the pellets and analyzed on Affymetrix microarrays. 7 biological replicates of both 3xMyc::FOG-1 and wildtype were used. FOG-3 RNA-coimmunoprecipitation (RIP) samples were prepared as follows. Parallel RIPs were performed with wild type animals as the control IP. Worm extracts were made from wildtype and FOG-3::3xFLAG late L3/early L4 animals. Myc affinity gel was used to immunoprecipitate FOG-3::3xFLAG from the extracts. RNA was extracted from the pellets and analyzed on Affymetrix microarrays. 7 biological replicates of both FOG-3::3xFLAG and wildtype were used.
Project description:Heart development is modulated by FOG-2/NuRD interactions. FOG-2R3K5A is unable to recuit the NuRD complex and this results in cardiac defects such as ASD, VSD, and thin ventricular walls We used microarrays to detail the changes in gene expression in FOG-2R3K5A hearts to determine misexpression of genes that may be causing the observed phenotypes.
Project description:We report ChIP-Seq data for GATA-1 and the FOG-binding mutant of GATA-1 (GATA-1^V205G) in G1ME cells, a Gata1-null cell line with both erythroid and megakaryocytic differentiation potential. We introduced HA-tagged GATA-1 or V205G into G1ME cells via retroviral transduction. The cells were crosslinked at 48h post-transduction, and an HA antibody was used for chromatin immunoprecipitation (ChIP). ChIP and input samples were sequenced on Illumina GAII high-throughput sequencer. The data reveal GATA-1-specific and V205G-specific bidning sites, indicating that FOG-1 both faacilitates and prohibits GATA-1 chromatin occupancy in a context-dependent manner. Examinaton of chromatin occupancy of GATA-1 anda FOG-binding mutant of GATA-1 in G1ME cells cultured in TPO.
Project description:In this experiment, steady-state mRNA levels were determined for replicated samples of N2 (wild-type reference) and fog-2(q71) homozygous mutant C. elegans. All samples were adult XX animals, which for N2 are self-fertile hermaphrodites and for fog-2(q71) spermless hermaphrodites, i.e. true females. For the fog-2 mutant animals, only those that had mated with males, and were thus gravid, were picked for RNA isolation. This ensures that all comparisons are between similar, embryo-containing animals. The experiment was motivated by the role of FOG-2 in post-transcriptional control of gene expression in germ cells, inferred from its the germline-specific phenotype of its loss and from its physical associated with the GLD-1 RNA-binding protein. Specifically, a possible role for FOG-2 in influencing mRNA stability was addressed.
Project description:We explored the role of FOG-1 in GATA-1 transcriptional regulation of megakaryocyte differentiation through expression of wild-type GATA-1 and the FOG-binding mutant of GATA-1 (GATA-1^V205G) in G1ME cells.
Project description:Rice plants cultivated on mountain areas are frequently exposed to acid fog and natural fog events. In this report, we analyzed the expression profiles of the rice plant with acid fog (SiAF) or neutral fog (SiNF) treatment for 1 and 7 days. Microarray results suggested that ROS generation was induced by not only SiAF, but also SiNF treatment, and it occurred in apoplast, not in organelles. Genes for defense- and stress-responses was also induced by both SiAF and SiNF treatments. The induction occurred in plants treated with SiAF for both 1 day and 7 days, and it was also detected in plants treated with SiAF for 7 days. These results suggest that both SiAF and SiNF treatments are abiotic stresses accompanying ROS generation in apoplast.
Project description:We explored the role of FOG-1 in GATA-1 transcriptional regulation of megakaryocyte differentiation through expression of wild-type GATA-1 and the FOG-binding mutant of GATA-1 (GATA-1^V205G) in G1ME cells. G1ME cells were derived from Gata1-null mouse ES cells and have both megakaryocyte and erythrocyte differentiation potential upon reconstitution of GATA-1 expression (Stachura 2006). HA-tagged wild-type or mutant GATA-1 were expressed in G1ME cells grown in TPO via retroviral transductions. The cells were sorted for GFP positivity 68 hours post-transduction and then were allowed to recover in normal growth medium for 4h. Total RNA was then isolated using RNeasy kit from Qiagen 72 hours post-transduction.
Project description:Heart development is modulated by FOG-2/NuRD interactions. FOG-2R3K5A is unable to recuit the NuRD complex and this results in cardiac defects such as ASD, VSD, and thin ventricular walls We used microarrays to detail the changes in gene expression in FOG-2R3K5A hearts to determine misexpression of genes that may be causing the observed phenotypes. Whole hearts from E16.5 Wild-type and FOG-2R3K5A siblings were processed for RNA extraction and hybridization on Affymetrix microarrays. We used three biological replicates of each genotype.