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. 2007 Mar 29;104(15):6329–6334. doi: 10.1073/pnas.0611655104

Fig. 2.

Fig. 2.

NK activating receptors are required for the polarization of the MTOC and cytolytic granules, whereas LFA-1 mediates conjugate formation. (A) YTS cells were conjugated with 721.221 cells in the presence of control mIgG or anti-CD28 mAb. The conjugates were fixed and permeabilized for intracellular staining of F-actin (green), microtubules (MT, red), and granules (blue) simultaneously. (B) The percentage of conjugates showing F-actin polymerization in the NKIS and MTOC and granule polarization to the NKIS (means and SDs). At least 50 conjugates images were examined in each case. The FACS-based conjugation assay was used with YTS-GFP cells and 721.221-RFP cells preincubated with control mIgG, anti-LFA-1 mAb, or anti-CD28 mAb. Results from six independent conjugation assays were averaged. (C) Similar imaging experiments were done with NK92 cells conjugated with K562 targets in the presence of either control mIgG, anti-LFA-1 mAb, or anti-NKG2D mAb. (D) The results of observing >50 images are shown with SDs. ∗, Percent F-actin polymerization and MTOC and granule polarization in the presence of anti-LFA-1 mAb are the values obtained in examining the small number of conjugates formed under these conditions.