Project description:The purpose of the experiment was to better understand the effects of antimigratory drugs lithium chloride and 6-bromoindirubin-3'-oxime (BIO) in glioblastoma cells. U251 cells were grown to 60-70% confluency in DMEM containing 10% FBS in 6 well plates. Drugs were added (LiCl 20 mM, BIO 5 micromolar, with carrier controls; either NaCl or DMSO) and incubated for 24 hours. Cells were then harvested and RNA purified using TRIZOL.
Project description:Bio-electrospray, the direct jet-based cell handling apporach, is able to handle a wide range of cells. Studies at the genomic, genetic, and the physiological level have shown that, post-treatment, cellular integrity is unperturbed and a high percentage (>70%, compared to control) of cells remain viable. Although, these results are impressive, it may be argued that cell based systems are oversimplistic. This study utilizing a well characterised multicellular model organism, the non-parasitic nematode Caenorhabditis elegans. Nematodes were subjected to bio-electrosprays to demonstrate that bio-electrosprays can be safely applied to nematodes.
Project description:MIXL1-GFP reporter lines (HESC3MIXL1-GFP/w and MEL1MIXL1-GFP/w, referred to as HES3 and MEL1 in this submission) were differentiated as spin embryonic bodies (Ebs) supplemented with mesodermal inducing growth factors (BMP4,SCF, VEGF based). Methylcellulose hematopoietic colony forming assays were performed by culturing dissociated day 4 EBs in the presence of hematopoietic growth factors (VEGF, SCF, IL3, IL6, TPO, EPO and FLT3L) for 7-9 days. Addition of WNT3a to the methylcellulose lead to the formation of compact, mesodermal colonies, which we term 'mesosphere' (Meso-balls). We demonstrated that the inclusion of WNT3a in the methylcellulose could be replaced with BIO, a GSK3 inhibitor, acting as a canonical WNT signaling agonist. Mesospheres formed in the culture supplemented with BIO molecules were termed 'BIO-balls'. Expression profiling between 'meso-balls' and 'BIO-balls' were compared and analyzed for markers assisiting in defining their phenotypes.
Project description:Human keratinocytes isolated from foreskin were cultured and treated with a GSK-3beta inhibitor, BIO. The effect of BIO was evaluated by the cell growth, clony formation and differentiating markers. We used microarrays to detail the global program of gene expression underlying cellularisation and identified distinct classes of up-regulated genes during this process.
Project description:Carboxy-terminally tagged MOZ (Flag-V5-BIO tagged) was detected by ChIP-seq using anti-V5 antibody (Sigma, A7345) to precipitate chromatin associated with MOZ
Project description:Human keratinocytes isolated from foreskin were cultured and treated with a GSK-3beta inhibitor, BIO. The effect of BIO was evaluated by the cell growth, clony formation and differentiating markers. We used microarrays to detail the global program of gene expression underlying cellularisation and identified distinct classes of up-regulated genes during this process. Human primary keratinocytes were subjected for RNA extraction. The total RNAs were hybridized on Affymetrix microarrays. The cultured cells exhibit a homogous cell morphology at passage 1 to passage 5. The cells were treated with BIO at passage 3 or 4.
Project description:To explore molecular mechanisms of different seed cells in bio-root regeneration, RNA sequencing (RNA-seq) was performed on adipose-derived stromal/stem cells (ASCs) and two dental derived stem cells