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Colocalization of VWF and FH in the Weibel-Palade bodies of ECs. HUVEC were grown on coverslips, fixed with PFA (4%), permeabilized with Triton X-100 (0.5%), and stained for intracellular VWF and FH. FH was stained with a pool of 3 biotinylated anti-FH antibodies: FH1 and FH2 (Quidel), and OX24 (AbDSerotec) followed by streptavidin-FITC. VWF was stained with a rabbit polyclonal anti-human VWF IgG (Dako) or a rabbit IgG control, followed by an Alexa Fluor 555-labeled goat anti-rabbit IgG. Nuclei were stained with Hoechst 33342 dye. Cells were washed and mounted in fluorescent mounting medium Fluoprep (bioMérieux). Images were acquired using an Axiovert 200M microscope (Zeiss) equipped with Apotome (63×). The image depicts the overlay of the green, red, and blue fluorescence. See the article by Rayes et al on page 121.