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. 2006 Jan 23;203(1):189–201. doi: 10.1084/jem.20050433

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Copyright © 2006, The Rockefeller University Press

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Figure 7. — (A and B) MLS_COX-GFP-GR is exclusively localized to the mitochondria. The mitochondria-directed GFP-GR variant was prepared by inserting the MLS of COX upstream to GFP-GR. HeLa cells (A), H1299 (B), or PC-3 (B) cells were transfected with either GFP-GR or MLS_COX-GFP-GR and stained with red mitrotracker to visualize the mitochondria before confocal microscopy. (C) MLS_COX-GFP-GR is more efficient in inducing apoptosis than GFP-GR. HeLa, L929 E8.2 A3, and PC-3 cells were transfected with plasmids encoding MLS_COX-GFP-GR or GFP-GR. After 48 h, the percentage of apoptotic GFP-positive cells (transfectants) was compared with that of GFP-negative cells (nontransfectants) of the same sample. (D) NLS-defective GRK513-515A induces apoptosis of HeLa cells. Cells were transfected with plasmids encoding GFP-GRwt, GFP-GRK513-515A, or pEGFP-F. After 48 h, the percentage of apoptotic cells was determined as in C. (E and F) A nucleus-directed GFP-GR variant (NLS_TAg-GFP-GR) induces apoptosis of HeLa (E) and H1299 (F) cells. Cells were transfected with plasmids encoding GFP-GR, MLS_COX-GFP-GR, NLS_TAg-GFP-GR, or pEGFP-F. After 48 h, the percentage of apoptotic cells was assessed as in C.