A. STATEMENT OF OBJECTIVES The Cell Biology Core is a new facility and will serve all four projects. This core will provide several lipoprotein-producing and non-lipoprotein producing cell lines required for studies described in each project. The lipoprotein producing cells will be used: 1) To study the mechanisms of assembly and secretion of endogenous apoA-l- and apoB-containing nascent lipoprotein particles;2) To determine the effects of several apoA-l mimetic peptides on the concentration, composition, and properties of apoA-l- and apoB-containing nascent lipoprotein particles;3) For transfection studies to assess the structure-function relationship of apoB, and potentially apoA-l, and their respective particle assembly and secretion;4) To silence the gene of interest using miRNA technique;5) To establish clonal stable transformants of cells that express C-terminally truncated forms of human apoB or cells that lack specific gene, i.e, lipid transfer proteins. The non-lipoprotein producing cell lines will be used: 1) For the expression of ABCA1, ABCG1, and ABCG4;2) To established clonal statble cell lines overexpressing ABCA1, ABCG1, and ABCG4;3) To evaluate the effects of apoA-l mimetic peptides on LPS-induced adhesion of THP-1 monocytes to HUVECs;4) For cholesterol efflux studies.
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