INTRODUCTION 2.1 The mucosal immune system 2.2 The inflammatory bowel disease 2.3 The effects of glucocorticoids 2.4 The mechanism of glucocorticoid receptor action 2.5 Immunsuppressive effects of glucocorticoids, the transrepression of NF-κB 2.6 Influence of cytokines on GR expression and function 3. MATERIALS AND METHODS 3.1 Materials 3.1.1 Laboratory devices 3.1.2 Tools 3.1.3 Cells 3.1.4 Chemicals 3.1.5 Solutions 3.1.6 Kits 3.1.7 Primers 3.1.8 Plasmid vectors 3.2 Methods 3.2.1 Cell culture

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... nditions 3.2.2 Protein isolation for Western blot II 3.2.3 Western blot 3.2.4 Immunostaining procedures 3.2.5 Confocal laser scanning microscopy (CLSM) 3.2.6 Transformation of E. coli 3.2.7 Purification of plasmid DNA 3.2.8 Transient transfection of Caco-2 and IEC-6 cells 3.2.9 SEAP Reporter Gene Assay 3.2.10 3 H-dexamethasone binding assay 3.2.11 RNA isolation 3.2.12 RT-PCR RESULTS Glucocorticoid receptor expression in Caco-2 and IEC-6 4.2 The gene regulatory effects of glucocorticoids can be measured by transfection experiments in intestinal cells 4.3 The transrepressive effect of dexamethasone on IL-1β-induced, NF-κB-mediated reporter gene activity in Caco-2 and IEC-6 4.4 IL-1β repressed the stimulating effect of dexamethasone on transactivation in both Caco-2 and IEC-6 cells 4.5 IL-1β inhibits glucocorticoid receptor translocation 4.6 IL-1β does not reduce dexamethasone binding in Caco-2 and IEC-6 4.7 Dexamethasone can reduce the IL-1β-induced TNFα expression only in IEC-6 cells but not in Caco-2 cells 4.8 Glucocorticoid receptor action could be restored by increasing the receptor content by GR-overexpression in Caco-2 4.9 NF-κB is not responsible for the impairment of glucocorticoid receptor action in Caco-2