ABSTRACT: Primary lung fibroblasts were grown to confluence in DMEM 10% FBS, serum starved in DMEM 0.5% FBS for 18 hours. Cells were then treated in the presence or absence of 100 nM ET-1 in the same medium for an additional 4 hours. Keywords: repeat sample
Project description:Primary lung fibroblasts were grown to confluence in DMEM 10% FBS, serum starved in DMEM 0.5% FBS for 18 hours. Cells were then treated in the presence or absence of 100 nM ET-1 in the same medium for an additional 4 hours.
Project description:primary lung fibroblasts p3 from normal periphery of resected cancer. grown in DMEM 10% FBS, until confluence, then DMEM 0.5% FBS for 18 hours. Media were then changed and incubated for an additional 4 hours in the presence or absence of 100 nM endothelin-1. Data represent two independent experiments performed on two different occasions. Keywords: other
Project description:primary lung fibroblasts p3 from normal periphery of resected cancer. grown in DMEM 10% FBS, until confluence, then DMEM 0.5% FBS for 18 hours. Media were then changed and incubated for an additional 4 hours in the presence or absence of 100 nM endothelin-1. Data represent two independent experiments performed on two different occasions.
Project description:LV fibroblasts were isolated from control (day 0), and infarcted regions at day 1, day 3, and day 7 after myocardial infarction (MI). Cells were isolated in DMEM in 10% fetal bovine serum and were seeded into 6-well plates and were allowed to grow to 80% confluence in DMEM supplemented with 10% fetal bovine serum (FBS). Cells were serum starved (DMEM + 0.1% FBS) for 18 h and then media was changed and secretome was collected after 24 h. The sample sizes are n=3 biological replicates for each time point.
Project description:We investigated how varying the composition of cell culture formulations and growing cancer cells at different densities might affect tumor cells genotype. Specifically, we compared gene expression profiles generated by human MDA-MB-231 human breast cancer cells cultured in different media (MEM, DMEM, or RPMI 1640) containing different concentrations of fetal bovine serum (FBS) or different sera (equine or bovine) that were grown at different cell densities. We have 6 different group (1. MDA-MB-231; MEM-10% FBS-90% confluence (MEM-1 and MEM-2). 2. MDA-MB-231; DMEM-10% FBS-90% confluence (DMEM-1 and DMEM-2). 3. MDA-MB-231; RPMI1640-10% FBS-90% confluence (RPMI-1 and RPMI-2) 4. MDA-MB-231; MEM-10% Horse serum-90% confluence (Horse-1 and Horse-2). 5. MDA-MB-231; MEM-0.1% FBS-90% confluence (FBS0.1-1 and FBS0.1-2). 6. MDA-MB-231; MEM-10% FBS-50% confluence (Con50-1 and Con50-2). Each group has duplicated. We analyzed; 1. Cell density (90% vs 50% confluence). 2. FBS concentration (10% FBS vs 0.1% FBS). 3. Sera (10% FBS vs 10% Horse serum). 4. Media (MEM vs DMEM vs RPMI1640)
Project description:3T3-L1 fibroblasts were cultured in high glucose DMEM supplemented with 10% foetal bovine serum (FBS) and 2 mM glutamax at 37°C in 10% CO2. For differentiation into adipocytes, fibroblasts were seeded into 12-well plates, and cultured in 1 mL/well of 10% FBS-supplemented DMEM containing an adipogenic cocktail (350 nM insulin, 0.5 mM 3-isobutyl-1-methylxanthine, and 250 nM dexamethasone) for 3 d, followed by 3 d in DMEM containing 10% FBS and 350 nM insulin. On day 9 post-differentiation, media volumes were either kept at 1 mL (high) or switched to 0.33 mL (low) for 16 h. Cells were then lysed in 350 µL RLT buffer for RNA extraction and sequencing.
Project description:A clonal doxycycline inducible dCas9-KRAB MDA-MB-231 cell line to control repression of CDK genes and PRMT5. On day 1 of the experiment cells were infected with lentiviruses containing the appropriate targeting/NTC sgRNAs driven by the human U6 promoter at an MOI of ~3 for each virus to ensure all cells were transduced. Cells were transduced in DMEM + 10% FBS with the addition of 8 μg/mL polybrene. 16 hours after the time of transduction, media was changed to DMEM +10% FBS. 24 hours after this, the cell culture media was switched to DMEM + 10%FBS containing 2μg/mL puromycin to ensure no uninfected cells remain. 48 hours after this, cell culture media was changed to DMEM + 10%FBS containing 2 μg/mL puromycin and 1 μg/mL doxycycline to induce dCas9-KRAB expression. 48 hours after this, cells were processed for CUT&Tag library prep following the manufacturer's recommendations.
Project description:Smad3 is an important downstream transcriptional factor of TGF-β signaling in the pathogenesis of renal fibrosis. Understanding the target genes is essential to decode the mechanism underlying Smad3-regulated renal fibrosis. To uncover the potential target genes of Smad3 in renal tubular cells, we applied the ChIP-seq technology to analyze the genomic chromatin binding landscape of Smad3 in TCMK-1 cells. The TCMK-1 cells of 90% confluence were starved in low-serum (0.5% FBS) medium for 6 h followed by stimulation with 5 ng/ml TGF-β1 for 30 min. The cells were harvested for ChIP assay. The precipitated chromatin was used for DNA isolation and deep-sequencing.