Project description:Angiogenesis is a highly orchestrated process involving complex crosstalk between several endothelial cell (EC) processes including cell cycle, cell survival and migration. Transcription factors ETS1 and ETS2 are required for EC functions necessary for embryonic angiogenesis. However owing to the lethal nature of the double mutant embryo, the specific gene targets of these factors are yet to be identified. In the current study, we try to elucidate the effect of endothelial cell specific deletion of Ets1 and Ets2 on post natal angiogenesis and characterize the downstream regulatory pathways. Deletion of Ets1 and Ets2, restricted to endothelial cells in new born mice (P1-P3), reduced retinal angiogenesis. Similarly, EC infiltration and invasion into matrigel plugs subsided when matrigel admixed with mouse mammary tumor cells was injected into adult mice with inactivated Ets1 and Ets2 specifically in ECs. Expression of key cell cycle and cell survival regulators were diminished in double mutant cells as compared to controls. In addition, both these factors were found to occupy the enhancer regions of the target genes. Deletion of Ets1 and Ets2 in cultured aortic EC resulted in altered cell cycle phases with a G2/M phase arrest and increased sensitivity to apoptosis in vitro. These results demonstrate that deletion of Ets1 and Ets2 in endothelial cells inhibits angiogenesis by altering cell cycle progression and decreasing cell survival.
Project description:Angiogenesis is a highly orchestrated process involving complex crosstalk between several endothelial cell (EC) processes including cell cycle, cell survival and migration. Transcription factors ETS1 and ETS2 are required for EC functions necessary for embryonic angiogenesis. However owing to the lethal nature of the double mutant embryo, the specific gene targets of these factors are yet to be identified. In the current study, we try to elucidate the effect of endothelial cell specific deletion of Ets1 and Ets2 on post natal angiogenesis and characterize the downstream regulatory pathways. Deletion of Ets1 and Ets2, restricted to endothelial cells in new born mice (P1-P3), reduced retinal angiogenesis. Similarly, EC infiltration and invasion into matrigel plugs subsided when matrigel admixed with mouse mammary tumor cells was injected into adult mice with inactivated Ets1 and Ets2 specifically in ECs. Expression of key cell cycle and cell survival regulators were diminished in double mutant cells as compared to controls. In addition, both these factors were found to occupy the enhancer regions of the target genes. Deletion of Ets1 and Ets2 in cultured aortic EC resulted in altered cell cycle phases with a G2/M phase arrest and increased sensitivity to apoptosis in vitro. These results demonstrate that deletion of Ets1 and Ets2 in endothelial cells inhibits angiogenesis by altering cell cycle progression and decreasing cell survival.
Project description:Expression profiling of Rag2-deficient Ets1++ and Rag2-deficient Ets1-- mature NK cells and WT bone marrow progenitors, WT T cells, and WT Pro B cells WT Hematopoietic progenitors, CD4 T cells, Pro B cells, and WT and Ets1-deficient NK cells were FACs sorted. RNA was subsequently extracted, labelled, and hybridized to Affymetrix microarrays. The goal if this experiment was to identify Ets1 dependent genes in NK cells
Project description:Expression profiling of Rag2-deficient Ets1++ and Rag2-deficient Ets1-- mature NK cells and WT bone marrow progenitors, WT T cells, and WT Pro B cells
Project description:RNA seq result shows that WT-AG-haESCs and DKO-AG-haESCs samples are clustered together using hierarchical cluster both in the all expression genes and imprinting genes. This suggests that DKO of DMRs of H19 and Gtl2 do not change the overall gene expression patterns in AG-haESCs.
Project description:To characterize the effect of loss of Ets1 in Non-TFH and TFH cells, we performed gene expression RNAseq analysis for T follicular helper (TFH) and Non-T follicular helper (Non-TFH) cells in WT (Ets1 fl/fl) and Ets1 KO (CD4-cre Ets1 fl/fl) mice.
Project description:We performed microarray analysis of gene expression in WT and Ets1-/- CD4+ CD8+ DP thymocytes. Overall, we find that Ets1-/- thymocytes display gene expression signatures closer to previous stages of thymocyte development (e.g. DN3-4) than WT DP cells, suggesting that while these cells do become DP thymocytes in the absence of Ets1, that the latter is required for the upregulation of later T-cell genes and that its presence is required for the downregulation of genes corresponding to earlier and alternative stages of development.
Project description:KMT2C and KMT2D are two of the frequently mutated epigenetic modifiers in urothelial cancer. To compare the histone post-translational modification with Kmt2c/d loss, we performed mass spectrometry analysis of histones from Kmt2c/d WT and dKO urothelial cells.