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. 2013 Sep 2;202(5):779–792. doi: 10.1083/jcb.201301004

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Figure 6. — Effect of dynein inhibition on MTOC repositioning. (A) Shown are the percent contributions of the four classes of MTOC repositioning scored 10 min after conjugation with +SEE Raji cells for Jurkats treated with CMP-2 (Inactive Analogue), Ciliobrevin-D (Cilio D), control siRNA (CTRL siRNA), siRNA against the dynein heavy chain (DHC siRNA), or overexpressing a dominant-negative fragment of p150 glued (CC1 DN). (B) Same as in A, except scored 30 min after conjugation. (C) Rates of MTOC movement during the polarization and docking phases. (D) MTOC-to-IS distance 10 min (untreated [UT] and Inactive Analogue) and 30 min (Cilio D, CTRL siRNA, DHC siRNA, CC1-DN) after conjugation. (E) Time-lapse montage showing MTOC repositioning in a representative conjugate containing a Jurkat cell treated with Ciliobrevin-D (merge of the transmitted image and the RFP-Pericentrin–labeled MTOC pseudocolored green). (F) As in E, except the Jurkat has been transfected with DHC-siRNA and the MTOC was labeled with GFP-Cen-2. (G) As in F, except the Jurkat was overexpressing RFP-CC1-DN. The yellow lines in E–G mark the IS membrane. The white arrowheads in E point to pauses and reversals in MTOC movement. The values for DHC siRNA and CC1-DN versus Cilo-D, and for CTRL siRNA versus CMP-2, were not statistically different. All data were obtained from at least three independent experiments.