Project description:Abl kinases, which are required for multiple cellular processes, including proliferation, apoptosis, adhesion, cell migration, and stress responses, widely participate in the regulation of gene transcription. To illustrate the role of Abl kinase in gene expression, the transcription of approximately 22,000 genes in wild-type (WT) and c-abl/arg knockout MEFs was detected using Affymetrix GeneChips (Mouse Genome 430 2.0).
Project description:mRNA profiles of beta-actin knockout (KO) and wild-type (WT) MEFs induced to adipocyte-like feature to study how global transcriptome changes during differentiation
Project description:This project looks at the effect a chemical modulator targeting PI3Kα has on the phosphoproteome of PI3Kα-WT and PI3Kα-KO MEFs. PI3K signalling is a critical regulator of numerous cellular processes such as proliferation, apoptosis, migration, invasion, metabolism, cell growth and autophagy. The PI3K pathway is frequently hyperactivated in cancer, commonly due to activating mutations or via loss of the lipid phosphatases that negatively regulate the pathway. We have recently identified a new small molecule modulator of the PI3K pathway and have used an unbiased phosphoproteomics approach to examine the impact of this compound on signalling pathways in cells that are wild-type and knockout for PI3K. Immortalised mouse embryonic fibroblasts (MEFs) isolated from PI3Kα-WT and PI3Kα-KO embryos were treated with and without the modulator (5 µM), exploring early (15 min) and late (4 h) time points. Insulin is a well-characterised activator of PI3K. Therefore, PI3Kα-WT MEFs were also treated with insulin (100 nM) as a positive control to compare with the PI3K modulator.
Project description:To gain insight into the signaling pathway(s) required for ABL1/ABL2-dependent bone metastasis, we evaluated the consequences of single or double inactivation of ABL1 and ABL2 on the transcriptome of breast cancer cells. Double ABL1/ABL2 knockdown was required to decrease the levels of p-CrKL by more than 90%, indicative of inactivation of the endogenous ABL kinases. To examine the consequences of depleting the ABL kinases on the transcriptome of metastatic breast cancer cells we employed next generation sequencing (RNAseq) analysis. We found that 180 genes were significantly down-regulated and 40 genes were significantly up-regulated in ABL1/ABL2 knockdown cells. Four samples were analyzed control, Abl single knockdown, Arg single knockdown, Abl/Arg double knockdown. Experiments were performed in triplicate.
