Abstract
59 Introduction 60 Results 63 ALCAM expression correlates with cell motility in uveal melanoma cell lines 63 Establishment of ALCAM-silenced MUM-2B cell lines by shRNA knockdown 65 ALCAM-silenced cells display reduced motility and invasive capacity 66 ALCAM overexpression is not sufficient to enhance migration and invasive capacity in MUM-2C cells 67 ALCAM-silenced cells exhibit reduced activation of MMP-2 67 Cadherin-based junctions are disorganized in ALCAM-silenced cells 68 ALCAM expression enhances cadherin-mediated cell-cell contacts in the 2C-ALC cell line 70 Discussion 70 CHAPTER FOUR: IDENTIFICATION OF NOVEL ALCAM EXTRAAND INTRACELLULAR LIGANDS 88 Abstract 88 Introduction 89 L1 cell adhesion molecule, an ALCAM extracellular interactor 89 Intracellular interactors 91 Results 91 L1 and ALCAM are expressed in the developing visual pathway 91 Evidence for a trans-interaction between ALCAM and L1 95 ALCAM and L1 interact in trans to regulate retinotopic mapping .........96 Isolation of eight candidate intracellular interactors by affinity chromatography 97 IQGAP1 97 Filamin A 98 α-actinin-4 99 Final comments 99 CHAPTER FIVE: CONLCUSIONS AND FUTURE DIRECTIONS 106 88 Introduction 89 L1 cell adhesion molecule, an ALCAM extracellular interactor 89 Intracellular interactors 91 Results 91 L1 and ALCAM are expressed in the developing visual pathway 91 Evidence for a trans-interaction between ALCAM and L1 95 ALCAM and L1 interact in trans to regulate retinotopic mapping .........96 Isolation of eight candidate intracellular interactors by affinity chromatography 97 IQGAP1 97 Filamin A 98 α-actinin-4 99 Final comments 99 CHAPTER FIVE: CONLCUSIONS AND FUTURE DIRECTIONS 106 The role of ALCAM in regulating N-cadherin-based adherens junctions ...107 IQGAP1 regulation of adherens junctions 110 Evidence for IQGAP1/ALCAM crosstalk 113 Filamin A regulation of transmembrane proteins at the cell surface .....117 Evidence for crosstalk between filamin A and ALCAM 118 α-actinin-4 regulation of the cadherin-catenin axis 120
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