Intestinal genetic inactivation of caspase-8 diminishes migration of enterocytes

Figure 1 Migration of caspase-8 (Casp8)-deleted Caco2 cells is diminished. A: Western blots Casp8 protein and densitometric analysis after treatment of Caco2 cells with small interfering RNAs (siRNAs) interfering with Casp8 transcripts or with negative control siRNA (∆scramble); B: Measurement of cellular proliferation by proliferating cell nuclear antigen (PCNA) expression in Caco2 cells with siRNA interfering with Casp8 transcripts or with negative control siRNA; C: Migration of Casp8 siRNA-treated Caco2 cells and scramble controls at 6 h and 24 h after scratching; D-F: Control siRNA-treated Caco2 cells at 0 h (D), 6 h (E), or 24 h (F) after scratching; G-I: Casp8 siRNA-treated Caco2 cells at 0 h (G), 6 h (H) or 24 h (I) after scratching.

Figure 2 Intestinal phenotype of Casp8^∆int mice. Morphologic pattern of A: Small intestinal; B: Large intestinal architecture in Casp8^f/f mice; C: Small intestinal; D: Large intestinal architecture in Casp8^+/∆int mice; E: Small intestinal; F: Large intestinal architecture in Casp8^∆int mice. Paneth cells are marked by arrows (hematoxylin and eosin,100 × magnification). Casp8: Caspase-8.

Figure 3 Morphometry examples of small and large intestinal tissues. Small or large intestinal tissue sections of Casp8^f/f, Casp8^+/∆int, and Casp8^∆int mice were anti-Ki67 (left) or anti-BrdU (right) immunostained. BrdU was injected 2 h, 20 h, or 40 h before sacrificing mice. Examples of cell^max locations in the small or large intestine are highlighted with a red dot (200 × magnification). Casp8: Caspase-8.

Figure 4 Tissue-specific analysis of enterocyte migration. Cell^max locations (enterocyte migration) in six histoanatomically distinct intestinal mucosal regions in cre-negative Casp8^f/f (f/f), heterozygous Casp8^+/∆int (+/∆int), and homozygous-knockout mice Casp8^∆int (∆int) at 2 h, 20 h, or 40 h after BrdU injection.