Figure 1. αII-Spectrin immunoblot, a positive control before HTPI.

Pooled naïve rat hippocampal lysate, lysates digested with calpain-2 or caspase-3 in vitro, and rat TBI hippocampal lysate. Separate naïve and TBI hippocampi from 6 individual animals respectively were pooled. To confirm that the extent of proteolysis in the last 3 samples was comparable, we analysed 20 μg of protein from each of these samples by traditional SDS/6% PAGE, immunoblotting and probing with monoclonal anti-αII-spectrin antibody (Affiniti anti-fodrin). Intact protein (280 kDa) was observed under all conditions. Calpain-2 digestion produced major fragments SBDP150 (150 kDa) and SBDP145 (145 kDa) (solid arrows), whereas caspase-3 digestion produced SBP149 (SBDP150i, 149 kDa) and SBDP120 (120 kDa) (open arrow heads) [5]. In TBI samples, a mixture of SBDP150, SBDP149, SBDP145 and SBDP120 was observed. M, molecular mass marker.