lec 28b - Lecture 28b Example of experiment using various...

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Lecture 28b: Example of experiment using various methods: siRNA, cell fractionation, Western blot and immunofluorescence Role of RanBP3 in β catenin nuclear export What happens when RanBP3 is depleted?
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nucleus cytoplasm plasma membrane β catenin Role of RanBP3 in β catenin nuclear export
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Wnt (= hormone) > stimulates intracellular signal > activated β catenin nucleus ‘activated’ β catenin (detected by specific antibody) stimulated cell, signal ON Signal OFF gene regulation in nucleus Regulation of the Wnt pathway Role of RanBP3 in β catenin nuclear export cytoplasm …….D S GIH S GAT T TAP S LSGK………… PO 4 PO 4 PO 4 PO 4 ‘inactive” Nterm …….D S GIH S GAT T TAP S LSGK………… ‘activated” Anti ”active” antibody
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Hendriksen et al, Journal of Cell Biology, 2005 Why only part of activated β catenin in nucleus? reducing nuclear export accumulation in the nucleus Hypothesis: maybe re exported? Prediction: Role of RanBP3 in β catenin nuclear export
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