DHC RNA interference prevents MTOC translocation. Jurkat cells (A) and CH7C17 cells (B). (left) The effect of DHC siRNA expression (siDHC). DHC was detected by Western blotting with anti-DHC goat polyclonal antibody. (middle) In control cells (siNeg), DHC is localized at the periphery of the IS, surrounding the MTOC. This accumulation does not occur in DHC-interfered cells (siDHC), which fail to translocate their MTOC. Conjugates were formed as in Fig. 1. DHC was detected with anti-DHC clone 440, followed by Alexa 568–labeled anti–mouse secondary antibody (red in the merge image). Tubulin was detected as in Fig. 1 (green). SEE-pulsed Raji and HA or SEB-pulsed Hom2 APC are blue in the merged image (CMAC-labeled). Differential interference contrast images are shown on the right. Bars, 10 μm. (far right) Quantification of MTOC translocation in conjugated Jurkat (A) or CH7C17 (B) T cells. More than 200 conjugates were counted for each condition. Data are means ± SD. *, P < 0.05 versus no stimulation; **, P < 0.001 versus siNeg (Student's t test).