Expression data fom human capillary network-derived cells before and after adipogenic differentation, and after chronic adenylate cyclase activation of differentiated cells
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ABSTRACT: Progenitors in human vasculature expanded in-vitro were differentiated with adipogenic cocktail for 12 days, following which they were stimulated with forskolin for 7 days Microarrays were used to detail the program of differentiation of thermogenic human adipose cells Adipose tissue from three individuals was cultured in hydrogel under pro-angiogenic conditions. Capillary networks emerging from the explants were subcultured on plastic, grown to confluence, subjected to adipogenic differentiation and after 12 days treated with vehicle or forskolin for 7 days
Project description:Progenitors in human vasculature expanded in-vitro were differentiated with adipogenic cocktail for 12 days, following which they were stimulated with forskolin for 7 days Microarrays were used to detail the program of differentiation of thermogenic human adipose cells
Project description:Progenitors in human vasculature expanded in-vitro were differentiated with adipogenic cocktail for 12 days, following which they were stimulated with forskolin for 7 days Microarrays were used to detail the program of differentiation of thermogenic human adipose cells
Project description:In order to gain insight into the molecular events which underlie auditory hair cell regeneration in chicken, we compared gene expression in chicken basilar papillae after 24, 48, and 72 hours in culture with or without forskolin (100uM).<br><br>Under sterile conditions, cochlear ducts containing the basilar papillae were carefully dissected out of ~4-day-old chicks and then individually cultured in DMEM with 10% fetal bovine serum with or without forskolin (final concentration 100 ?M, delivered in 1% DMSO) for either 24, 48, or 72 hours at 37ï¾°C. Control samples received DMSO at 1% as a vehicle control. At the end of 3 days, the tegmentum vasculosum was dissected off to expose the auditory epithelium, which was delicately freed from the underlying cartilaginous plates. All explants were kept in culture for 3 days because new hair cells are first seen in basilar papillae treated with forskolin after ~3 days of exposure to the drug [19]. Therefore, at even earlier time points in culture the molecular events that subserve hair cell proliferation are well underway. Each sample was comprised of 3 auditory epithelia from 3 different chicks which were put in ~100 ?L of DMEM and then immediately frozen at -80ï¾°C until RNA isolation could be performed. There were a total of 24 samples in this experiment: six 72-hour forskolin, six 72-hour control, three 48-hour forskolin, three 48-hour control, three 24-hour forskolin, and three 24-hour control.
Project description:PC12 cells (passage 22) were cultured at 5% CO2 in a 37 C incubator. The growth medium was DMEM (high glucose, Invitrogen) supplemented with 25mM HEPES, 10% FCS, 5% FBS and 1x Pen/Strep (Invitrogen). 1x10^6 cells were starved overnight in DMEM +25 mM HEPES and treated with 10 uM 1,9 dideoxy forskolin or forskolin for 60 min. The final DMSO concentration was 0.05%. Total RNA was purified using Trizol (Invitrogen) according to the manufacturer's protocol. An additional round of purification was conducted using Rneasy columns (Qiagen) according to the manufacturer's protocol. cRNA was synthesized and labeled according to standard Affymetrix protocols. 2 biological replicates for the forskolin and 1,9 dideoxy forskolin conditions were run on Affymetrix RAE230 A and B chips for a total of 8 hybridizations.
Project description:Long term exposure to incretin hormones is known to have salutory effects on beta cell function and viability. While short-term cAMP induction is known to have a signature CREB-CRTC target gene response, the long-term effects of cAMP on beta cell gene expression are less well understood. We used rat microarray analysis to compare the genome-wide gene expression response to short-term (2 hours) and long-term (16 hours) stimulations of the cAMP agonist forskolin in INS-1 insulinoma cells. INS-1 cells were exposed to forskolin for 2 or 16 hours in RPMI medium containing 10% serum. Control samples wer incubated for the same time without forskolin. Extracted RNA was used for hybridization on Affymetrix rat 1.0 st gene arrays.
Project description:In this study we showed that rat XEN cells grown in the presence of a GSK3 inhibitor exhibited enhanced formation of cell contacts and decreased motility. In contrast, treatment with forskolin induced the PE formation and epithelial-mesenchymal transition (EMT) in rat XEN cells. Using microarray and real-time PCR assays, we found that VE versus PE formation of rat XEN cells was correlated with change in expression levels of VE or PE marker genes. Similar to forskolin, EMT was prompted upon treatment of rat XEN cells with recombinant parathyroid hormone related peptide (PTHRP), an activator of the cAMP pathway in vivo. Taken together, our data suggest that rat XEN cells are PrE-like cells. The activation of Wnt pathway in rat XEN cells leads to the acquisition of VE characteristics, whereas the activation of the PTHRP/cAMP pathway leads to EMT and the formation of PE. Rat XEN cells were cultured in four different conditions with 3 parallels for each condition: (1-3) without treatments control, (4-6) 2 days treated with CHIR alone, (7-9) 1 day treated with CHIR and 1 day with both CHIR and forskolin, and (10-12) 1 day treated with forskolin alone. In all samples on DAY1 cells were plated, at DAY2 cells were cultured in usual conditions (DMEM F12 medium and 10% Fetal Bovine Serum, both from Sigma) and inhibitor of GSK3 kinase 3 μM CHIR99021 (Axon Medchem) was added in samples 4-9. On DAY3 all cells were cultured in medium with 0.1% serum to exclude the influence of serum and in CONTROL (1-3) cells were cultured without experimental treatments, in CHIR (4-6) cells were further cultured with 3 μM CHIR99021, in CHIR plus FORSKOLIN cells were cultured with 3 μM CHIR99021 and 10μM Forskolin (Sigma), in FORSKOLIN cells were cultured for 1 day with 10 μM Forskolin. On DAY4 RNA was isolated.
Project description:miRNA profiling of mouse kidney arteriolar smooth muscle cells (aSMCs) of the renin lineage comparing control untreated cells with cells treated with forskolin to induce renin expression. Two condition experiment: control untreated aSMCs vs forskolin treated aSMCs; Biological replicates: control 3, treated 3; independently grown and harvested. One replicate per array.
Project description:The experiment was performed to identify PKA phosphorylation substrates in wildtype and autophagy-deficient brains. Therefore, hippocampus and cortex of wildtype and conditional knockout mice were isolated and sliced. The brain slices were incubated with DMSO or Forskolin, a cAMP elevating agent, to induce PKA activity. Samples were measured by LC-MS/MS and were used to quantify proteomic and phosphoproteomic changes.
Project description:Expression profiling of HEK293T cells transfected with miR-neg (negative control), miR-132 or miR-381 mimics and treated with forskolin for 0, 1 or 4hrs. HEK293T cells were reverse transfected with the respective miRNA mimics. 72hrs post-transfection, cells were treated with forskolin for 0, 1 or 4hrs. Total RNA was then extracted. Expression profiling was carried out with Affymetrix arrays. Experiment was performed in duplicate. Total of 18 samples.