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Bodipy FL LDL endocytosis in HepG2 cells
HEPG2 cells were plated onto poly-D-lysine-coated glass bottom dishes in growth medium and allowed to recover for 24 hours before switching to starvation medium overnight, using Fluorobrite DMEM medium (Cat. No. A1896702) plus 0.3% BSA. Bodipy LDL (Cat. No. L3483) (pseudocolor green) was added to the medium at 10 µg/mL and the cells were returned to the incubator for 3 hours. NucBlue Live nuclear stain (Cat. No. R37605) (pseudocolor blue) and LysoTracker Deep Red (Cat. No. L12492) (pseudocolor red) was added to the cultures at 1X concentration at 37°C in last 30 minutes. Cells were then washed twice in Assay Buffer solution (HBSS plus 3% BSA) and the images were captured on the EVOS FL Auto 2 Imaging System. Data shown was overlaid with white light, GFP light cube for Bodipy LDL (pseudocolor Green), Cy5 light cube for Lysotracker Deep Red (pseudocolor red), and DAPI light cube for NucBlue Live stain (pseudocolor blue).
