FIGURE 7.

Hsc70 expression affects the subcellular distribution of TH in MN9D cells. A, an enriched SV preparation was prepared from WT, scramble shRNA, and Hsc70 shRNA cells by differential centrifugation. 50 μg of total protein from the original homogenate (H) as well as the supernatants (S) and pellets (P) from each step were analyzed by SDS-PAGE and IB using TH, SYPH, and Hsc70 antibodies. SYPH expression was enriched in the P3 sample compared with H and S3 of all three cell lines, confirming that the P3 sample represented enriched SVs. Consistent with our previous studies, TH expression was found in P3 of WT and scr-shRNA cells. Interestingly, this TH localization is shifted from P3 to S3 in Hsc70 shRNA cells. B, specific TH activity was examined in the S3 and P3 fractions of MN9D WT cells (left panel), MN9D cells expressing the scr-shRNA (center panels), and MN9D cells expressing the Hsc70 shRNA (left panels). *, p < 0.05.