Figure 2

Living cells expressing fusion proteins and immunostained fixed cells. Representative fluorescent images of living U-2 OS cells expressing endoG-EYFP (green) and AMID-tHcRed (red) (A), AMID-tHcRed (B), and AMID-tHcRed 6 hours after induction of apoptosis by addition of 200 nM staurosporine (C). Representative fluorescent images of stably transfected living cell U-2 OS over-expressing endoG-EYFP (D), living cell U-2 OS expressing endoG-EYFP before (E) and 6 hours after (F) apoptotic induction by 200 nM staurosporine. Fixed U-OS cells immunostained with fluorescently labeled primary antibodies against human AIF (red) and cyclophilin A (green) (G), U-2 OS cell 6 hours after induction of apoptosis by 200 nM staurosporine immunostained against human AIF (H) and cyclophilin A (I). Representative fluorescent images of fixed U-2 OS cells immunostained with fluorescently labeled primary antibodies against human DNA topoisomerase II α at non-apoptotic condition (J), 3 (K) and 6 (L) hours after induction of apoptosis by 200 nM staurosporine. Representative fluorescent images of fixed U-2 OS cells immunostained with fluorescently labeled primary antibodies against human HSP70-1 after heat shock for 2 hours at 42°C (M), fixed U-2OS cells immunostained against human HSP70-1 before (N) and 6 hours after (O) apoptotic induction by 200 nM staurosporine. Bottom parts of figures J, K, L, N, O contain representative sections of XZ projections of acquired 3D images. Scale bars = 10 μm.