Comparison of the Redox Potential in the Endosomes of Different Cell Types By Imaging Live Cells Via FRET Microscopy
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Sub-cellular compartments have different reduction-oxidation (redox) environments and these electrochemical differences play a vital role in their distinct functions. Although much research has been conducted on the mitrochondria, nuclei, secretory pathway, and extracellular space, much remains to be elucidated regarding the redox potential in the endocytic pathway. Additionally, although some cells have been studied in this regard such as macrophages and dendritic cells, there exists no real comparison across different cell as to whether there is any variance in the redox potential across the endocytic pathway. In this study fluorescence resonance energy transfer (FRET)-based redox-sensitive fusion protein probes were used to compare FRET signals across cancer cells (PC3 and BT549 cell line), fibroblasts, and bone marrow-derived dendritic cells in order to draw conclusions about the redox potential in the endocytic pathways. The results show that the redox-sensitive reporter encapsulated inside pH-sensitive liposomes could be taken up by the different cells and tracked while alive by using FRET. These results suggest that endosomes have a varied reducing environment that depends on the cell tested. In comparing the redox data with previously obtained Bone Marrow Macrophage (BMMO) data, these preliminary results show that all the cells tested were less reducing than the BMMOs though statistical differences have not been established. These findings may have implications for drug delivery via the endocytic pathway across different types of cells.