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. 2002 Mar 1;22(5):1738–1751. doi: 10.1523/JNEUROSCI.22-05-01738.2002

Fig. 3.

Fig. 3.

Dose- and time-dependent accumulation of cytosolic cytochrome C in MMT-treated PC12 cells. A, Western blot.B, Cytochrome C ELISA assay. A, Subconfluent cultures of undifferentiated PC12 cells were harvested at 1 and 3 hr after treatment with 200 or 500 μm MMT. The cytosolic fractions were obtained as described in Materials and Methods. Cytosolic fractions were separated by 12% SDS-PAGE, transferred to a nitrocellulose membrane, and cytochrome C (Cyt C) was detected using polyclonal antibody raised against cytochrome C. For β-actin measurements, the membrane used for cytochrome C was reprobed with β-actin antibody to confirm equal protein loading in each lane. The immunoblots were visualized using ECL detection agents from Amersham. B, Subconfluent cultures of undifferentiated PC12 cells were harvested at 15 and 30 min after treatment with 200 or 500 μm MMT. The cytosolic fractions were obtained as described in Materials and Methods. The value of each treatment group is the mean ± SEM from two separate experiments in triplicate. Asterisks (*p < 0.05) indicate significant differences compared with vehicle-treated cells.