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. 2001 Oct 1;21(19):7439–7446. doi: 10.1523/JNEUROSCI.21-19-07439.2001

Fig. 6.

Fig. 6.

Time course of procaspase-3 protein expression in rat brain cortex after TBI. A, Fifty microgram aliquots of cytosolic protein extracts isolated from sham control or traumatized rat cortex at indicated times after TBI were subjected to 12% SDS-PAGE and transferred to a nitrocellulose filter. The filter was probed with a rabbit polyclonal antibody against caspase-3 (Casp-3; H-277; Santa Cruz Biotechnology). The antigen–antibody complexes were visualized by an ECL method as described in Materials and Methods. To control protein loading, membranes were stripped and reprobed with an antibody against β-actin. B, Fifty microgram aliquots of cytosolic protein extracts isolated from sham control or traumatized rat cortex at indicated times after TBI were incubated with or without active recombinant human caspase-9 (20 U; Biomol, Plymouth Meeting, PA) in 50 μl of caspase activation buffer at 37°C for 1 hr. Caspase-3-like activity was assayed fluorometrically by measuring the accumulation of free AMC. Protease activity is expressed as a percentage of the activity in sham-operated control extracts.