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bis_104_pq_ldls_fall_08 - OBSERVATION When exogenous LDLs...

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OBSERVATION : When exogenous LDLs are added to cells in culture the LDLs appear to bind to the plasma membrane in discrete clusters which are then internalized as clathrin coated pits. Hypothesis #1: Prior to exposure to LDLs, LDL receptors (LDL-Rs) are randomly and diffusely distributed around the cell’s PM. Binding of LDLs induces the lateral movement and clustering of the ligand-receptor complexes into clathrin coated pits prior to endocytosis. Hypothesis #2: Cells from individuals with the FH -/- mutation have normal amounts of LDL-Rs in their PMs, but they are incapable of lateral movement and fail to form clusters upon LDL binding. Design a series of experiments to test the plausibility and validity of these hypotheses. Expt a: Immunostain live cells from a normal individual with a fluorochrome tagged antibody against the external domain of the LDL-R. Apply this antibody to two batches of cells, before and after treating them with LDLs. Compare the fluorescent staining patterns by fluorescence microscopy.
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