Fig. 6. Translocation of metallacycle 2. (A) Subcellular localization of metallacycle 2 (red) in HeLa cells by staining with the special fluorescent tracers, lysotracker (green) and Hoechst 33342 (blue). Metallacycle 2 localization analysis (the first column); z-stack mode analysis (the second column); three-dimensional (3D) reconstruction and co-localization analysis (the third column). (B) Intracellular uptake of the metallacycle in HeLa cells after pretreatment with different endocytosis inhibitors. HeLa cells were pretreated with various endocytosis inhibitors for 1 h and then incubated with 10 μM of metallacycle for different periods of time at 37 °C or 4 °C. The control group was incubated with metallacycle 2 only at 37 °C. (C) Changes in the UV-Vis spectrum of metallacycle 2versus time in PBS buffer at pH = 5.3 containing lysozymes (c = 10 μM).