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Cells were labeled with Lysotracker Red DND-99 (Invitrogen) for 2 h at 1000× dilution or 20 μg/mL DQ Green BSA (Invitrogen) overnight in mESC medium before fixing with 4% paraformaldehyde for 20 min at 25 °C. Cells were then stained with Hoechst stain. To induce starvation, cells were cultured in serum free media for 16 h before harvesting. Images were acquired with fluorescence microscopy (Cell Observer, Zeiss).
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