Figure 3

Bcl-2 expression and cytochrome c release in cultured neurons and astrocytes. Western blotting revealed that low dose minocycline upregulated Bcl-2 expression of neurons (A: lysates from 0, 1 and 100 μM of minocycline-treated neurons correspond to lanes 1, 2, and 3, respectively) with subsequent inhibition cytochrome c release from mitochondria to cytosol (B: lysates from 0, 1 and 100 μM of minocycline-treated neurons correspond to lanes 7, 8, and 9, respectively). In contrast, minocycline at all doses did not upregulate Bcl-2 expression in astrocytes (A: lysates from 0, 1 and 100 μM of minocycline-treated astrocytes correspond to lanes 4, 5, and 6, respectively). Immunocytochemical analysis revealed that minocycline low dose (D: 1 μM) significantly increased the number of Bcl-2 positive neurons compared to vehicle-treated (C: 0 μM) or high dose-treated neurons (E: 100 μM). In contrast, minocycline at all doses did not alter the number of Bcl-2 positive astrocytes (F: 0 μM, G: 1 μM and H: 100 μM). Quantitative analyses of Bcl-2 positive cells are shown in panels I and J. Data represent mean values ± SEM (* p < 0.05. I: neurons, J: astrocytes). Scale bar: 25 μm.