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. 2000 May 1;149(3):613–622. doi: 10.1083/jcb.149.3.613

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© 2000 The Rockefeller University Press

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Reduction of apoptotic cell numbers after ischemic brain injury in caspase-11^−/− mice. A, Western blot of spleen lysate from 3 wild-type (+/+) and 4 caspase-1 (−/−) mice, probed by anticaspase-11 (top), anticaspase-1 (middle) and antitubulin (bottom). L929 cell lysate (L929) was used as a positive control. B, TUNEL (a, c, and e) and Hoechst dye (b, d, and f) staining of wild-type (a, b, e, and f) and caspase-11^−/− (c and d) brains. a–d are from ischemic brains and e and f are from untreated control brain. Permanent ischemia was induced by occlusion of the left middle cerebral artery using monofilament as described by Hara et al. 1997a. 12 h after the occlusion, brains were processed for TUNEL. C, Quantification of TUNEL-positive cells in the wild-type and caspase-11^−/− ischemic brain. The percentages of TUNEL-positive cells were determined by the number of TUNEL-positive cells divided by the number of Hoechst dye-positive cells. TUNEL- and Hoechst dye-positive cells were quantified using the Zeiss Axiovert microscope equipped with Northern exposure software. Mean of the reading from five brains is shown. Error bar represents standard error.

Reduction of apoptotic cell numbers after ischemic brain injury in caspase-11^−/− mice. A, Western blot of spleen lysate from 3 wild-type (+/+) and 4 caspase-1 (−/−) mice, probed by anticaspase-11 (top), anticaspase-1 (middle) and antitubulin (bottom). L929 cell lysate (L929) was used as a positive control. B, TUNEL (a, c, and e) and Hoechst dye (b, d, and f) staining of wild-type (a, b, e, and f) and caspase-11^−/− (c and d) brains. a–d are from ischemic brains and e and f are from untreated control brain. Permanent ischemia was induced by occlusion of the left middle cerebral artery using monofilament as described by Hara et al. 1997a. 12 h after the occlusion, brains were processed for TUNEL. C, Quantification of TUNEL-positive cells in the wild-type and caspase-11^−/− ischemic brain. The percentages of TUNEL-positive cells were determined by the number of TUNEL-positive cells divided by the number of Hoechst dye-positive cells. TUNEL- and Hoechst dye-positive cells were quantified using the Zeiss Axiovert microscope equipped with Northern exposure software. Mean of the reading from five brains is shown. Error bar represents standard error.