Fig. 3. Maintenance of stress fibers by DAP kinase does not require its death domain. (A) NIH3T3 cells were co-transfected with vector for wild-type DAP kinase or DAPKDD and the GFP expression vector at a ratio of 10:1. Cells were serum starved for 6 hours, lysed and subjected to western blot analysis. (B) Alternatively, serum-starved transfectants were double stained with rhodamine-phalloidin and Hoechst 33258. Transfected cells are visualized by GFP fluorescence. Scale bar, 10 µm. (C) Quantitation of GFP-positive cells with stress fibers in experiments as described in (B). (D) NIH3T3 cells were transiently transfected with control vector, DAP kinase or DAPKDD. Two days after transfection, cells were re-fed with serum-free medium and then cultured for another 18 hours. Apoptotic cells were determined by cell death detection ELISA as described previously (Wang et al., 2002).

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