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. 2017 Apr 25;112(8):1703–1713. doi: 10.1016/j.bpj.2017.01.038

Figure 5.

Figure 5

Image correlation of the plasma membrane by TIRF-SIM and STICS analysis in Jurkat T cell forming immunological synapses after α-actinin CRISPR knockout. (a) Image and quantification of α-actinin knockout were normalized to wild-type Jurkat T cells; Western blots show both anti-ACTN1 and anti-GAPDH as a control. (b) The plasma membrane was labeled with the lipophilic dye DiO, and imaged 5 min after contact with an antibody-coated coverslip. Scale bars, 5 μm. (c) STICS output vector maps are shown. (d) Plots of the cell means for membrane flow velocity (2.8 ± 0.67 μm) and membrane flow directionality (80.4 ± 19.7°) are shown. Histograms show the normalized (e) speed within the dSMAC and (f) directionality. n = 16 (controls) and 8 (α-actinin−/−); n.s., nonsignificant; ∗∗p < 0.005. To see this figure in color, go online.