Figure 9

Effect of calpain and caspase-3 inhibitors on glutamate-induced cleavage of α-fodrin in primary cortical cells. Primary cortical cells isolated from 18 day old fetal rat brains were cultured for 7–8 days, and then cells were treated with 1 mM glutamate alone or in the presence of calpain inhibitor 12.5 μM ALLN and caspase-3 inhibitor 50 μM z-DEVD for 6 hrs. (I): representative Western blots showing calpain and caspase-3- mediated proteolysis of α-fodrin and inhibition by protease inhibitors. (II): semi-quantitative analysis of calpain and caspase-3-mediated proteolysis of α-fodrin. Accumulation of breakdown products was assessed by Western blot analysis as described under "Methods". Actin was used as a loading control. The bars depict densitometric analysis of Western blots from at least three experiments (± SEM) *P < 0.05 compared to control; † P < 0.05 compared to glutamate alone.