Figure 3.

Gating Strategy Illustration

Intracellular staining of FVIII on U937 cells, Zenon labeling. (A–C) Logical gating strategies are shown on (A) forward light scatter (FSC)/side light scatter (SSC) to define the populations and to exclude (B) doublet cells and (C) death cells/debris. (D and E) FVIII positivity (E) is then compared to the appropriate IgG isotype (D). Superimposed plots and histograms include the following: unstained cells (black) and cells stained with only fixable live marker (light gray) superimposed to FVIII-positive cells (blue) and relative IgG (light blue). (F–H) Different domains in HeLa and U937 cells (F and G) are visualized with different intensities (H) (mean ± SD, n = 2 on cells after FVIII transduction).