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. 2006 Apr 17;203(4):973–984. doi: 10.1084/jem.20050625

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Copyright © 2006, The Rockefeller University Press

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Figure 1. — Characterization of primary IEC preparations. (A) Visualization of isolated primary epithelial cell aggregates comprising intestinal villi with the attached crypts by phase contrast microscopy. Bar, 1 μm. Phalloidin staining of freshly isolated primary cells illustrating actin accumulation along apical microvilli. Bar, 30 μm. (B) Verification of the polarized phenotype of primary IECs by immunostaining for E-cadherin (left) and ZO-1 (right). The depicted images show a three-dimensional reconstruction of the polarized, multicellular epithelial cell complex to illustrate the cellular structure. Bar, 10 μm. (C) Determination of cell viability and apoptosis of primary IECs by propidium iodine (PI) exclusion analysis and FITC–annexin V staining. (D) Staining for nonepithelial hematopoietic (CD45⁺) cells of a representative preparation of primary IECs using anti-CD11b, anti-CD3, and anti-B220 antibodies.