Analysis of the expression and cellular distribution of ADTRP, a novel protein that coregulates with TFPI. (A) Coexpression plot (red) of ADTRP and TFPI resulted from the GAMMA of GEO microarrays shows significant positive correlation between the expression levels (represented as fold-change) of the 2 genes. P < .0001 for the slope's deviation from zero. n = 1500. Confidence interval (CI; 95%) is shown as black interrupted lines. (B) Western blot performed on (i) cellular lysates of HEK293T (positive control, lane CTR) and EAhy926 (lane EC), and (ii) tissue extracts of normal baboon lung and human placenta, using rabbit anti-ADTRP and anti-TFPI IgGs for probing. (C-F) Immunofluorescence and confocal microscopy show the localization of ADTRP in human ECs in culture and the baboon aorta. Non-permeabilized HUVECs (C) were immunostained for cell surface ADTRP (FITC, green) and TFPI (Cy₃, red). EA.hy926 cells (D) were first immunostained for cell surface ADTRP (FITC, green) and TFPI (Cy₃, red), then permeabilized and immunolabeled for Cav-1 (Cy₅, blue). Overlap (white) designates the colocalization channels obtained by Adobe Photoshop image analysis. Baboon aorta sections were immunostained for ADTRP (FITC, green) and Cav-1 (E) or TFPI (F; Cy₃, red for both) after brief permeabilization.^10,11  Blue: nuclear staining. Bars: (C-D) 10 μm; (E-F) 50 μm.