Project description:Analysis genes which are elevated after miR10a inhibitor transfection for 24 hours with transfection efficiency more than 95 %inhibition in mesangial cells. The hypothesis is genes which up-regulated significantly can be potential targets for miR-10a in primary human mesangial cells.
Project description:we performed genome-wide transcriptome profiling in mesangial cells and tubular epithelial cells (TECs), which were stimulated by high glucose (HG) and detected the expression of inflammation associated genes. HG increased the mRNA expression of oxidative stress, inflammasome and mammalian target of rapamycin (mTOR) related genes in mesangial cells. Pro-inflammatory/Th1 gene expression was upregulated, but Th2 related gene expression was downregulated in mesangial cells. In TECs, HG stimulation increased pro-inflammatory/Th1/Th2 gene expression. mRNA profiles of human mesangial cells and tubular epithelial cells, which were stimuated HG for 24 hours or mannitol contorol.
Project description:Quiescent human fibroblasts (2091) were transfected with 100nM mature miR-22 RNA duplexes. Cells were collected 24 hours after miR-22 transfection.
Project description:CRISPR KO RNA-seq data: To generate knockouts, we used a commercially available arrayed CRISPR-Cas9 library targeting 81 out of ~100 DUBs and 13 additional proteins in the ubiquitin-proteasome system, including ubiquitin-like proteins; the library was constructed with four, pooled, guides per target. mRNA profiling was performed 96 hours after guide RNA transfection using a high-throughput, low-cost RNA-sequencing method (3’ Digital Gene Expression or 3’DGE-seq) in the MDAMB231 breast cancer cell line. Inhibitor RNA-seq data: For the small molecule inhibitor signatures, MDAMB231 or MCF7 or MCF7 TP53 shRNA breast cancer cells were treated with a panel of DUB inhibitors for 24 hours, then 3'DGE-seq was used to perform the mRNA profiling.
Project description:Activation of GFI1-super-enhancer (GFI1-SE) by a LSD1 inhibitor NCD38 was relevant to myeloid differentiation and antileukemia effect in human erythroleukemia cells (HEL cells). Thus, we investigated the role of GFI1-SE upon NCD38 treatment in HEL cells. We established three independent sublines with bi-allelic deletion of GFI1-SE (CCE2 #114, #141, and #216) using CRISPR-Cas9 genome editing system in HEL cells and a classical limiting dilution method. Control sublines (Ctrl C1, C2, and C5) were established by transfection of parent vector and limiting dilution. After treatment of each subline with DMSO or NCD38 (LSD1i) for 24 hours, these gene expression profiling data were obtained.
Project description:Human primary keratinocytes were depleted of GRHL3 by siRNA and induced to differentiated for 2 days by addition of Calcium Primary normal human keratinocytes were transfected with GRHL3 or scrambled control siRNA using RNAi max (Life Technologies). 24 hours post transfection medium was raised to 1.8mM to induce differentiation. Cells were collected 48 hours later.
Project description:To investigate the molecular mechanism of the hypopigmentation observed in Dicer KO mice, Dicer knockdown was realised in vitro in normal C57BL/6 mouse melanocyte Melan-a cells. Transient transfection of Melan-a cells with siRNA directed against Dicer reduced Dicer protein levels to approximately 40% of that of siScr Melan-a cells 24 hours after transfection. We analyzed the transcriptome of Melan-a cells 24 hours after transfection with siDicer or siScr to understand the molecular mechanisms involved in Dicer-dependent melanocyte migration.
Project description:To investigate the molecular mechanism of the hypopigmentation observed in Dicer KO mice, Dicer knockdown was realised in vitro in normal C57BL/6 mouse melanocyte Melan-a cells. Transient transfection of Melan-a cells with siRNA directed against Dicer reduced Dicer protein levels to approximately 40% of that of siScr Melan-a cells 24 hours after transfection. We analyzed the miRnome of Melan-a cells 24 and 48 hours after transfection with siDicer or siScr to understand the molecular mechanisms involved in Dicer-dependent melanocyte migration.
Project description:Human primary keratinocytes were depleted of MLL2 by siRNA and induced to differentiated for 2 days by addition of Calcium Primary normal human keratinocytes were transfected with MLL2 or scrambled control siRNA using RNAi max (Life Technologies). 24 hours post transfection medium was raised to 1.8mM calcium to induce differentiation. Cells were collected 48 hours later.
Project description:Today, the pathogenesis of human enterovirus type 71 (HEV71) infection in human central neural system remains unclear. HEV71 is the major pathogen of hand-foot-and-mouth disease (HFMD), and has been associated with severe neurological disease and even death in infants and young children. We employed the human whole genome microarray analyze the mRNA profiling in human neuroblastoma cells SH-SY5Y infected with HEV71 after transfection. Firstly, SH-SY5Y cells were transfected wtih miR-1246 inhibitor and negtive control respectively using HiPerFect Transfection Reagent according to the manufacturer’s instructions. Then the cells were infected with HEV71 after transfection. After 12 hours infection, the cells were harvested to microarray analysis. The results showed the altered expression of mRNAs including up-regulated genes and down-regulated genes. Overall, this finding will help to understand the functional genes in HEV71-infected human neuroblastoma cells and miR-1246-virus-host interaction.
