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. 2001 Mar 19;193(6):755–768. doi: 10.1084/jem.193.6.755

Figure 1.

Figure 1

Figure 1

Figure 1

Efficient conditional ablation of VCAM-1 in vivo. (A) Southern blotting strategy for detection of IFN-α–induced VCAM-1 gene deletion. (B) Southern blot analysis of splenic DNA of mice without (lanes 1 and 2) and with (lanes 3 and 4) IFN-α injection (probed for VCAM-1). Without either the cre transgene (lanes 1 and 3) or IFN-α induction (lane 2), deletion of the VCAM-1 gene did not occur, as indicated by the 3- and 6-kb bands. By contrast, deletion is almost complete after IFN-α induction in the presence of the cre transgene (lane 4), resulting in a 4.5-kb signal. (C) Southern blotting of various tissues from IFN-α–treated, conditional VCAM-1 mutant mice revealed a high degree of deletion in most organs, but almost no deletion in brain (LI, liver; LN, lymph node; LU, lung; SP, spleen; TH, thymus; BR, brain; HE, heart; IN, intestine; KI, kidney). (D) Western blot analysis of VCAM-1 (110 kD) in conditional VCAM-1 mutant and control mice. After IFN-α–induced Cre expression, VCAM-1 could not be detected in BM, liver, lymph node, spleen, and thymus of conditional VCAM-1 mutant mice (lanes 4, 6, 8, 10, and 11), while minor traces of VCAM-1 are detectable in heart, intestine, and lung (lanes 2, 5, and 7) and brain contains VCAM-1 levels similar to control mice (lane 1). (E) Immunohistochemical analysis of spleen reveals VCAM-1 expression in control mice (left) and by contrast, no VCAM-1+ cells in spleen of conditional VCAM-1 mutant mice (right).