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. 2007 Oct 1;204(10):2305–2320. doi: 10.1084/jem.20061893

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Copyright © 2007, The Rockefeller University Press

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Figure 9. — CIP4 function in MTOC polarization relative to Cdc42 activation. The cytolytic IS between a KT86 cell and a YTS cell nucleofected with constitutively active Cdc42V12-GFP and GAPDH siRNA (A–F) or CIP4 siRNA (G–L) are shown using DIC microscopy (A and G), as well as confocal microscopy showing fluorescence for α-tubulin (B and H), pericentrin (C and I), CIP4 (D and J), Cdc42V12-GFP (E and K), and an overlay (F and L). (M) The mean percentage of the pericentrin-defined MTOC that colocalized with the tubulin-defined MTOC (P/M) and the percentage of the α-tubulin–defined MTOC that colocalized with the pericentrin-defined MTOC (M/P) in YTS cells nucleofected with Cdc42V12-GFP and GAPDH siRNA (black bars) or Cdc42V12-GFP and CIP4 siRNA (gray bars), and conjugated with KT86 target cells, are shown. (N) Mean distance in micrometers of the pericentrin-defined MTOC to the IS in YTS cells nucleofected with Cdc42V12-GFP and GAPDH siRNA, or Cdc42V12-GFP and CIP4 siRNA. The increase in cells receiving CIP4 siRNA was significant. *, P < 0.01. (O) Active Cdc42 pull down and Cdc42 Western blot from YTS cells activated with immobilized anti-CD28. Before activation, cells were nucleofected with GFP or Cdc42V12-GFP (top), or nucleofected with GAPDH siRNA or CIP4 siRNA (middle). Activated YTS cells were also compared with activated YTS cells overexpressing WT CIP4 (bottom). Error bars represent the SD.