Figure 6: Analysis of CD8⁺ T-cell polarization by IFC.

(a) CTV-labelled F5 CD8⁺ T cells were activated as indicated and expression of CD8 (upper row) and PKC-ζ (lower row) assessed with division. Location of the undivided peak (red dotted line) is identified relative to unstimulated controls (b) Representative multispectral, spatially registered 40 × images of late mitotic F5 CD8⁺ T cells in the absence (left panels) or presence (right panels) of cytochalasin B for the last 4 h of culture. Cells were stained with CD8 and PKCζ, as well as protein content and division history determined by CTV fluorescence. The plane of cytokinesis was established using the propidium iodide (PI) nuclear image (denoted by white dotted line). * Denotes example of asymmetric CD8 expression. (c,d) Polarity of each signal was calculated across daughter poles of 111 late mitotic cells entering first division and significance determined using a one-way analysis of variance.