Project description:Gene expression profiling of human embryonic kidney (HEK293) cells was performed to determine the effect of high and low glucose on gene expression. Microarrays were used to identify distinct classes of genes up-regulated in HEK293 cells during cultivation for 7 days in medium with physiological (low) glucose compared to high glucose. Human embryonic kidney cells (HEK293) were cultivated for 7 days in commercially available DMEM (supplemented with 10 % FCS) containing high glucose (450 mg/dl) or low glucose (100 mg/dl). Cells were harvested, total RNA was extracted and microarray gene expression profiling was performed to compare differential gene expression between low and high glucose conditions. Two biological replicates for each condition were made (high glucose: HEK-high-gluc-1 and HEK-high-gluc-2; and low glucose: HEK-low-gluc-1 and HEK-low-gluc-2).
Project description:Gene expression profiling of human embryonic kidney (HEK293) cells was performed to determine the effect of high and low glucose on gene expression. Microarrays were used to identify distinct classes of genes up-regulated in HEK293 cells during cultivation for 7 days in medium with physiological (low) glucose compared to high glucose.
Project description:We established a novel model to assess the function of proteins under in vivo conditions. The model relies on the expansion of HEK293 cells in immunodeficient NOD.Scid mice. To validate the novel model, we performed microarray gene expression profiling of NOD.Scid-expanded HEK293 cells relative to conventionally cultivated cells. Microarray analysis revealed that cell expansion in NOD.Scid mice restored an imbalanced chaperone system without inducing a major upregulation of the entire protein folding machinery. Human embryonic kidney (HEK293) cells were injected subcutaneously into immunodeficient NOD.Scid mice. After three weeks, the expanded cell pellet was isolated, and total RNA was extracted. In parallel, total RNA was prepared from HEK293 cells cultivated in vitro under standard cell culture conditions in a commercially available medium containing 450 mg/dl glucose (DMEM). Microarray gene expression profiling was performed to determine differential gene expression between in vivo expanded and in vitro cultivated HEK293 cells. Two biological replicates for each condition were made (in vitro cultivated HEK293 cells: Cells-1 and Cells-2; and NOD.Scid-expanded HEK293 cells: Scid-1 and Scid-2).
Project description:This gene array analysis shows that the transcriptome of HEK293, representing low-differentiated human embryonic kidney cells, cultured in presence of tRA and cAMP differentiating agents, is consistent with a genetic program relevant to kidney development
Project description:This gene array analysis shows that the transcriptome of HEK293, representing low-differentiated human embryonic kidney cells, cultured in presence of tRA and cAMP differentiating agents, is consistent with a genetic program relevant to kidney development Comparison of HEK293 cells grown in standard contitions or in presence of differentiating agents. Cells were seeded in 10 cm dishes, cultured for 3 days either in complete medium or in medium supplemented with tRA and cAMP, harvested, and spun down before RNA isolation. Comparative analysis allowed us to identify genes transcriptionally by tRA/cAMP.
Project description:Analysis of gene expression in human embryonic kidney cells HEK293 overexpressing the transcription factors HNF4a2 or mutant forms HNF4a2 C106R and HNF4a2 R154X, which were introduced by FRT/FLP recombination. The analysis was performed 24 hours following 1µg/ml tetracycline treatment to induce the expression of the genes. Results identify HNF4a regulated genes in kidney cells being several of these genes deregulated in renal cell carcinoma. Keywords = HEK293 embryonic kidney cells Keywords = HNF4a2 Keywords = tetracycline Keywords = FRT Keywords: ordered
Project description:To investigate the effects of the KDM4 inhibitor QC6352 on embryonic renal tumor cells, we treated WiT49 anaplastic Wilms tumor cells and tumor-forming HEK293 human embryonic kidney cells with 25 nM QC6352 for 72 hours
Project description:A time course of infection of the alphavirus Sindbis virus (SINV) was used to investigate the presence of viral specific vsRNA and the changes in miRNAs profiles in human embryonic kidney 293 cells (HEK293) by high throughput DNA sequencing. Deep sequencing of small RNAs early in SINV infection (4 and 6 hpi) showed low abundance (0.8%) of viral specific RNAs (vsRNAs) , with a random uniform distribution not typical of Dicer products, suggesting they arise from non-specific degradation. Sequencing showed little variation of cellular microRNAs (miRNAs) at 4 and 6 hpi compared to uninfected cells. Twelve miRNAs exhibiting some minor differential expression by sequencing, showed insignificant modulation by Northern blot analysis.
Project description:This model is from the article:
Competing G protein-coupled receptor kinases balance G protein and β-arrestin signaling
Heitzler D, Durand G, Gallay N, Rizk A, Ahn S, Kim J, Violin JD, Dupuy L, Gauthier C, Piketty V, Crépieux P, Poupon A, Clément F, Fages F, Lefkowitz RJ, Reiter E. Mol Syst Biol.
2012; 8: 590. 22735336
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Abstract:
Seven-transmembrane receptors (7TMRs) are involved in nearly all aspects of chemical communications and represent major drug targets. 7TMRs transmit their signals not only via heterotrimeric G proteins but also through β-arrestins, whose recruitment to the activated receptor is regulated by G protein-coupled receptor kinases (GRKs). In this paper, we combined experimental approaches with computational modeling to decipher the molecular mechanisms as well as the hidden dynamics governing extracellular signal-regulated kinase (ERK) activation by the angiotensin II type 1A receptor (AT(1A)R) in human embryonic kidney (HEK)293 cells. We built an abstracted ordinary differential equations (ODE)-based model that captured the available knowledge and experimental data. We inferred the unknown parameters by simultaneously fitting experimental data generated in both control and perturbed conditions. We demonstrate that, in addition to its well-established function in the desensitization of G-protein activation, GRK2 exerts a strong negative effect on β-arrestin-dependent signaling through its competition with GRK5 and 6 for receptor phosphorylation. Importantly, we experimentally confirmed the validity of this novel GRK2-dependent mechanism in both primary vascular smooth muscle cells naturally expressing the AT(1A)R, and HEK293 cells expressing other 7TMRs.