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Flow Cytometry And Biochemical Analysis Of Apoptotic Mouse Ht-2 T - Lymphocytes, Nell Pinkston
Flow Cytometry And Biochemical Analysis Of Apoptotic Mouse Ht-2 T - Lymphocytes, Nell Pinkston
Honors Theses
Apoptosis is a highly organized intracellular death program in multicellular animals. In the vertebrate immune system, apoptosis plays a central role in preventing the emergence of autoreactive lymphocytes. In this study, we used multiple stimuli (staurosporine, camptothecin, and cytokine deprivation - interleukin-2 or IL-2) to initiate apoptosis in IL-2 dependent, mouse HT-2 T-lymphocytes. All three inducers triggered DNA laddering and phosphatidylserine externalization. Propidium iodide staining and flow cytometry were also used to determine whether apoptotic cells accumulated in a specific stage of the cell cycle, and whether the mode of induction affected cell cycle distribution. Our findings indicate that IL-2 …
An Investigation Of Bag3 Knockdown And Its Effects On The Cytotoxicity Of Laromustine, Colin Sheehan
An Investigation Of Bag3 Knockdown And Its Effects On The Cytotoxicity Of Laromustine, Colin Sheehan
Honors Theses
Laromustine is a sulfonylhydrazine anticancer prodrug whose main cytotoxic arises from its ability to interfere with DNA replication of dividing cells. Multiple studies have suggested that Laromustine induces a form of cell death known as apoptosis. In a previous study investigating the mechanism of apoptosis, bcl2-associated athanogene 3 (BAG3) demonstrated significant upregulation in the presence of Laromustine. Given its anticancer ability, we selected BAG3 as a target for further investigation. BAG3 knockdown through transient shRNA transfections was performed in U138 glioblastoma multiforme cells and verified using qRT-PCR analysis. Finally, cell death assays were used to assess Laromustine’s cytotoxic effect on …