FIGURE 7.

Perturbations of Rab35 or EPI64C impair conjugate formation. A, quantitation of conjugate formation between transfected Jurkat cells and Raji cells in the presence or absence of superantigen SEE assessed by flow cytometry. Labels indicate the construct with which the Jurkat cells were transfected. Values plotted are the mean ± S.E. for results from five independent experiments. ^**, p < 0.015. B, representative images of Jurkat cells transfected with either dominant negative GFP-Rab35 (S22N) or constitutively active GFP-Rab35 (Q67L) in physical contact with Raji cells (stained with CellTracker Blue) with or without SEE. C, Jurkat cells were transfected with shEPI-A and shEPI-B vectors, and lysates were immunoblotted as indicated. Quantitation is shown of conjugate formation between Jurkat cells (transfected with the indicated construct) and Raji cells in the presence or absence of superantigen SEE assessed by flow cytometry. Values plotted are the mean ± S.E. for results from three independent experiments. ^**, p < 0.015. D, quantitative scoring of enrichment of TCR at the IS in 25 cells from RFP- and Rab35 DN-transfected Jurkat. Bars represent mean ± S.E. from three independent experiments. ^**, difference is significant at p = 0.01 by the nonparametric Mann-Whitney test.