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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 4. — Transient postnatal activation and LPS susceptibility of primary IECs. (A) Characterization of intracellular MIP-2 staining using nonactivated or LPS (100 ng/ml) -stimulated peritoneal macrophages. (B) Intracellular MIP-2 staining of primary IECs isolated from late gestational fetal, 6-, and 28-d-old mice. MIP-2 was cytometrically analyzed after 6 h of incubation in the presence of brefeldin A under nonstimulating conditions or after the addition of 100 ng/ml LPS. Dotted line, isotype control. (C) Double immunostaining and FACS analysis of primary IECs from 6-d-old mice with MIP-2 and E-cadherin to identify spontaneous MIP-2 production in IECs. (D) MIP-2 (red) in IECs from 6-d-old mice was detected by intracellular staining with a rabbit anti–MIP-2 antiserum (MIP-2) or an irrelevant affinity-purified rabbit control serum after incubation for 6 h in the presence of 0.5 μg/ml brefeldin A. Isolated IECs were identified as epithelial cells by E-cadherin staining (green). Counterstaining with DAPI. Bar, 20 μm.