Figure 5.

Erk1/2 de-activation kinetics are altered in ezrin-silenced cells. (A) Jurkat J77 cells transfected with siRNA control, or siRNA ezrin-1 were activated with soluble anti-CD3 Ab (MEM92) for the indicated times. Cells were then fixed, permeabilized, stained with anti-pErk1/2 Ab and analysed by flow cytometry. (B) Jurkat J77 cells were treated with 2 μg/ml of colchicine for 30 min, then activated and processed as in (A). Histograms represent fluorescence intensity of pErk versus number of cells. Graphs were rescaled to have the peaks at the same height using FlowJo software, to facilitate the comparison of curve shapes. The significance of the differences in cell distribution between activated and non-activated populations was calculated by comparing curve shapes as described in Supplementary data. ‘P'-value is indicated in each panel. Dashed lines mark the mean fluorescence intensity values of unstimulated or maximally activated control cells. The figure shows a representative experiment out of three independent experiments carried out. NS=non-significant.