DNA binding, expression, and cellular localization of SF1

A: Electromobility shift assay showing wild–type (WT) and mutant SF1 binding to a labeled probe corresponding to the 3′ SF1 binding site of the promoter. in-vitro translated empty vector (−) (lane 1) and an excess of unlabeled cold probe (with WT SF1 protein) (lane 7) were used as controls. B:Cellular localization of GFP–SF1 fusion proteins (green), generated and expressed in tsa 201 cells using a pAcGFP–C1 vector. Nuclear counterstaining was performed with DAPI (blue), and images merged to conform nuclear localization.WT SF1 showed strong nuclear localization, with relative nucleolar exclusion and very occasional nuclear subfoci. A similar expression and localization pattern to WT was seen for the p.R84H mutant. Clustering in larger subnuclear foci was seen in the cells transfected with the p.C33S mutant.The p.Y138 Xmutant showed di¡use nuclear localization.Copyright information:Taken from "Five novel mutations in steroidogenic factor 1 (SF1, ) in 46,XY patients with severe underandrogenization but without adrenal insufficiency"Human Mutation 2008;29(1):59-64.Published online Jan 2008PMCID:PMC2359628.Copyright © 2008 Wiley-Liss, Inc., A Wiley Company