Journal: Frontiers in Immunology

Article Title: Ezrin/Radixin/Moesin Proteins and Flotillins Cooperate to Promote Uropod Formation in T Cells

doi: 10.3389/fimmu.2013.00084

Figure Lengend Snippet: Impact of expression of ezrin constructs on random human T cell migration and chemotaxis . (A,B) T cells were transfected with EGFP or wild type ezrin or T567D ezrin or 1–310 ezrin as indicated and placed 4 h later on a layer of IL1β-stimulated human endothelial cells in the absence of added chemokine. Random migration of the transfected T cells was assessed for 10 min by time-lapse videomicroscopy at 37°C. The % of migrating transfected T cells (A,C) and the mean speed of the migrating transfected T cells (B,D) was evaluated. Mean ± SEM of three experiments. * P < 0.05, ** P < 0.01 for statistically significant differences between data as indicated (total number of T cells analyzed in three experiments: 247/656 cells transfected with EGFP; 376/574 cells transfected with wild type ezrin; 437 cells transfected with T567D ezrin; 679 cells transfected with 1–310 ezrin). (E) T cells were transfected with the indicated constructs and placed 4 h later into filter inserts. The lower and upper wells (open bars) or only the lower wells (closed bars) contained 200 ng/ml SDF-1. The number of transfected migrated T cells in the lower wells was determined after 1 h and calculated as % of total transfected T cells placed initially into the upper wells. In the absence of SDF-1 in the lower wells migration was zero for all samples and experiments. Mean ± SEM of eight experiments for SDF-1 only in the lower wells and of three experiments for SDF-1 in the upper and lower wells. * P < 0.05, *** P < 0.001 for statistically significant differences between data as indicated.

Article Snippet: 20 h prior to the experiment hCMEC/D3 cells were stimulated with 20 ng/ml recombinant interleukin-1β (IL1β) (PromoKine, Vitaris).

Techniques: Expressing, Construct, Migration, Chemotaxis Assay, Transfection