Project description:This experiment was investigating how gut commensal bacteria and intestinal inflammation affect miRNA expression. We analyzed miRNA expression of spleen and intestine from specific pathogen free (SPF) B6 mice, germ-free (GF) B6 mice, and IL-10 knockout mice which have severe colitis by microarray. Thus we have total 6 samples: GF spleen; GF intestines; SPF spleen; SPF intestine; IL-10 KO spleen and IL-10 KO intestine. We directly isolated RNA from whole spleens or intestines without any treatments, and then did microarray analysis.
Project description:OE17 and OE23 are chloroplast targeted proteins. RNA co-immunoprecipitation was performed using antibodies raised against these proteins. RNA from the pellet and from the supernatent for each pulldown was labeled with different fluoro-dyes.
Project description:Co-immunoprecipitation of the maize pentatricopeptide protein PPR4 and extraction of bound RNA. RNA is labelled and hybridized to a maize chloroplast genome tiling array in order to identify target RNA species of PPR4.
Project description:Identification of target transcripts for the putative chloroplast RNA binding protein CFM2 in Zea mays. CFM2 was immunoprecipitated from a chloroplast extract. Chloroplast extracts were prepared from WT tissue. RNA from the pellet and from the supernatant for each pulldown was labelled with different fluoro-dyes and hybridized onto an array covering the complete maize chloroplast genome. Messages enriched in the immunoprecipitate from WT tissue are likely targets for CFM2.
Project description:Mouse glomeruli and brain capillary fragments were prepared. Podocytes were separated from isolated glomeruli from 8-day-old Podocin-Cre, Z/EG double transgenic mice as follows: Isolated glomeruli were incubated with trypsin solution containing 0.2 % trypsin-EDTA (Sigma-Aldrich), 100ug/ml Heparin and 100U/ml DNase I in PBS for 25 min at 37 degree, with mixing by pipetting every 5 min. The trypsin was inactivated with soybeans trypsin inhibitor (Sigma-Aldrich) and the cell suspension sieved through a 30 um pore size filter (BD bioscience, Franklin Lakes, NJ). Cells were collected by centrifugation at 200 x g for 5 min at 4 degree and resuspended in 1ml PBS supplemented with 0.1 % BSA. To separate GFP-expressing (GFP+) and GFP-negative (GFP-) cells, glomerular cell were sorted using a FACSVantage SE (BD, San Jose, CA, USA) operating at a sheath pressure of 22 psi. Autofluorescent cells were excluded by analyzing the emission of orange light (585 nm). To allow for standardization of results, the samples such as glomerular RNA, brain capillary RNA, rest of kidnay RNA, podocyte RNA and foxc2 glomreular RNA were hybridized in competition with common reference samples.
Project description:Up-regulation of motility genes and biosynthesis genes were found in the pck expressing high ATP cell by transcriptome analysis while catabolism genes were down-regulated. Keywords: response depends on intracellular ATP concentration derived by pck or ppc overexpression 1. pck or ppc overexpressing E. coli at early log phase using glucose-minimal medium 2. pck or ppc overexpressing E. coli at chemostat culture (D=0.1 h-1)using LB-glucose medium
Project description:In order to take an unbiased approach and discover all the locations of Cse4 in the genome, we utilized formaldehyde crosslinking and immunoprecipitation followed by hybridization to DNA microarrays. We analyzed the location of Cse4 in three different strains; one in which the endogenous Cse4 is tagged with 12Myc epitopes (Cse4-12Myc), and one which contained both the endogenous Cse4 (untagged) and an ectopic copy of Cse4-12Myc expressed from the GAL1-10 promoter (pGAL1-10-Cse4-12Myc), and one in which Cse4 is tagged with 3HA epitopes (Cse4-3HA). ChIP-chip was performed using custom microarrays, to look at the genome-wide localization of the centromeric histone variant Cse4. Biological replicates were performed for each Cse4 epitope-tagged strain