H/I injury-induced pyramidal neuron death in the hippocampi of the wild-type and caspase-3-deficient neonatal mouse brains. A: Double staining of cleaved caspase-7 (Casp7) and DAPI in the contralateral hippocampus of a caspase-3-deficient mouse brain 8 hours after H/I injury. An arrow shows a cleaved caspase-7-positive neuron. B–E: Triple staining of cleaved caspase-3 (B, D) or cleaved caspase-7 (C, E), TUNEL, and DAPI in the ipsilateral hippocampi of wild-type (Wild) (B, C) and caspase-3-deficient (D, E) neonatal mouse brains 8 hours after H/I injury. F and G: The cleavage of caspase-7 (F) and GAPDH (G) in the contralateral (CL) and ipsilateral (IL) hippocampi of caspase-3-deficient mouse brains 8 hours after H/I injury. In the ipsilateral hippocampus of a caspase-3-deficient mouse brain, a cleaved form appears intensely at ∼27 kDa but not at 18 kDa (F, middle), whereas an activated form of caspase-7 can be detected at ∼18 kDa under a longer exposure (F, right). These cleaved forms, along with the proform of caspase-7, correspond with those detected in a control lane (Cont) showing primary hepatocytes treated with tumor necrosis factor-α and actinomycin D for 6 hours (F, left). Scale bars: 10 μm (A); 30 μm (E).