Overexpression of ADTRP increases TFPI expression and activity. (A) Immunofluorescence and confocal microscopy showing the distribution and colocalization of cell surface TFPI (Cy₃, red) with YFP-TF (YFP, green) and Cav-1 (Cy₅, blue; after permeabilization) on TF-EC stable expressing ADTRP-FLAG. Triple overlap (white) and nonoverlapping TF (green) channels result from the image analysis. See also Figure 3A (top panel) for control TF-EC images. Bars: 10 μm. (B) Scatter-plot and statistics (1-way ANOVA) of the colocalization between TF and TFPI, and TF/TFPI/Cav-1, out of the total MFI of TF. Data are mean ± SEM. ***P < .001 for both sets of data in the ADTRP-group compared with Control TF-EC group. ns, P ≥ .05 for the differences between sets of data within the 2 experimental groups. (C) The inhibitory capability of cell surface TFPI against TF-FVIIa–dependent FXa generation measured in TF-EC in the presence/absence of inhibitory anti-TFPI IgG. Values are normalized to 10⁶ cells. ***P < .001 versus control (unpaired t test). Data are mean ± SD of triplicates.