Development of a flow cytometric assay to study glucocorticoid receptor-mediated gene activation in living cells
作者： B Necela
DOI: 10.1016/S0039-128X(03)00032-1
关键词:
摘要: Abstract A flow cytometry-based reporter gene assay was developed and utilized to measure glucocorticoid receptor (GR)-mediated activation at the single cell level in living cells. generated that contains two copies of response element an E1b TATA box upstream a destabilized enhanced green fluorescent protein. Glucocorticoid Cos-1 HTC lines measured vivo by cytometry shown be dose dependent, leading increase total fluorescence population. Flow cytometric analysis indicated this per sample resulted from number cells expressing activated protein (GFP) as well overall mean GFP within Activation activity time dependent occurring early 1–2 h after dexamethasone addition. specific it exhibited different sensitivities range glucocorticoids could blocked with antagonists. Coexpression coactivator SRC-1a or P65 subunit NF-kappa B GR led enhancement repression, respectively. Taken together, these data suggest reporter-based is effective method for analyzing receptor-mediated expression
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