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. Author manuscript; available in PMC: 2010 Feb 1.
Published in final edited form as: Arch Biochem Biophys. 2008 Dec 10;482(1-2):58–65. doi: 10.1016/j.abb.2008.11.028

Figure 2. Intracellular distribution of TFPI-2 in endothelial cells assessed by immunoblotting and immunocytochemistry.

Figure 2

Figure 2

To investigate the intracellular distribution of endogenously expressed TFPI-2 protein in endothelial cells, HUVEC, HAEC and DMEC cells were cultured under standard growth conditions. (A), For immunoblotting, lysate and cell fractions from all cells were prepared and probed with anti-TFPI-2 IgG. The purity of cytosolic and nuclear fractions was verified using anti-alpha-tubulin and anti-histone-H1 antibodies, respectively. For immunoblot analysis, the intensity of blot bands were assessed by densitometric semi-quantitation and depicted by a bar diagram (lower panel). (B), DMEC cells were grown on two-chamber culture slides and immunostained with murine monoclonal antibody SK-9 and Alexa Fluor-555-conjugated goat anti-mouse IgG as the secondary antibody. The nucleus is counterstained using DAPI in mounting media. (a), vehicle plus Alexa Fluor-555-conjugated goat anti-mouse IgG. (b), SK-9 antibody and Alexa Fluor-555-conjugate goat anti-mouse IgG.