The effect of a dominant-negative RIP-K45A on TNF-stimulated L929 cellsPublic
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Tumor Necrosis Factor -alpha (TNF) is a regulatory cytokine of the immune system that acts on cells to induce apoptosis via recruitment of death-domain - interacting proteins. Receptor interacting protein (RIP) is an example of such a protein. It includes nuclear factor-KB, but triggers programmed necrosis when caspases are inactivated. The purpose of this project was to determine whether the mouse fibro sarcoma cell line L929 provides a valid model for studying TNF induced programmed necrosis. Indeed, treatment of L929 cells within TNF followed by measurement of cell viability with an MTS assay indicated that TNF induces cell programmed necrosis in these cells. Next the cells were transferred with a plasmid encoding a dominant-negative form of RIP to test the effects of inhibiting RIP activity on TNF signaling. Cell death (as measured by a propidium iodide assay) was not dramatically altered in RIP-inhibited cells. Thus, additional components may regulate programmed necrosis in L929 cells.
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