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

Figure 3.

Figure 3.

Colocalization of CIP4 with the MTOC and accumulated F-actin at the cytolytic IS. Cytolytic conjugates between a YTS GFP cell and a KT86 cell (A–F), and an NK92 cell and a K562 GFP cell (G–L). DIC images (A and G) and GFP expression in the YTS cell (B), or in the K562 cell (H), distinguish the NK cell from the target cell. Confocal microscopy for F-actin (C and I), α-tubulin (D and J), CIP4 (E and K), and an overlay (F and L) are shown. (M) Molecular accumulations in NK cells from ≥150 conjugates over at least three experiments between NK92 and K562 cells, YTS GFP and KT86 cells, YTS CIP4 and KT86 cells, CIP4 FCH− and KT86 cells, and CIP4 SH3− and KT86 cells. CIP4 accumulation was not determined for the CIP4-overexpressing cells, as it was diffuse. (N) The total area of F-actin, the tubulin-defined MTOC, and CIP4 measured in ≥15 conjugated NK cells over three experiments between NK92 and K562 cells (black bars), YTS GFP and KT86 cells (light gray bars), and YTS CIP4 and KT86 cells (dark gray bars). CIP4 mutants were not detected with anti-CIP4 mAb. (O) The percent colocalization among the areas in ≥15 conjugated NK cells over three experiments between NK92 and K562 cells (black bars), YTS GFP and KT86 cells (light gray bars), and YTS CIP4 and KT86 cells (dark gray bars) was calculated to demonstrate the percentages of the MTOC colocalized with CIP4 (M/C), of the CIP4 colocalized with the MTOC (C/M), of the F-actin colocalized with CIP4 (A/C), and of the CIP4 colocalized F-actin (C/A). Differences between YTS GFP and NK92 cells were not significant, but those between YTS CIP4 and YTS GFP cells were. Error bars represent the SD. *, P < 0.01.