Effects of 17β-E
and Δ8, 17β-E
on glutamate-induced Cyto c release into cytosol in primary cortical cells. Primary cortical cells isolated from 19 day old fetal rat brains were cultured for 7 days and treated with 1 mM glutamate alone or in the presence of 17β-E2 and Δ8, 17β-E2. After 6 h, cells were collected and cytosol fraction was prepared as described under "Methods". Cytosolic protein levels of Cyto c were determined by Western blot analysis as described under Figure 6. Actin bands were used to monitor the same blot to verify consistency of protein loading. The content of Cyto c was measured by densitometric scanning of the film. The results are the mean (± SEM) of at least three experiments. a = P < 0.05 compared with control; b = P < 0.05 compared with glutamate treated cells.