(A) Representative TIRF and dSTORM images of NKG2D on pNK cells incubated for 10 min on slides coated with PLL, MICA and ICAM-1, or ULBP2 and ICAM-1, blocked with specific antibodies against MICA or ULBP2 as appropriate, and then stained with fluorescently labelled specific mAb against NKG2D. Scale bars, 4 μm. Regions outlined in white are magnified and shown with corresponding density images according to the pseudocolor scale bar, thresholded binary maps, and Ripley’s K analysis. Scale bars, 1 μm. (B) Nanocluster areas (left), nanocluster density (middle), and percentage of localizations in nanoclusters (right) for NKG2D from the data shown in (A). Each symbol represents the median of several 5 μm x 5 μm regions from one cell. Lines represent means ± SD. Data are from a minimum of 12 cells from two independent donors. Each color represents one donor. *P < 0.05 and **P < 0.01 by one-way ANOVA with Tukey’s post-hoc test. Nonsignificant differences are not indicated.