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pHrodo Red 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. pHrodo Red LDL (Cat. No. L34356) (pseudocolor red) 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) was added to the cultures at 2 drops per mL 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, RFP light cube for pHrodo Red LDL (pseudocolor Red) and DAPI light cube for NucBlue Live stain (pseudocolor blue).
