HeLa cells were treated with 30uM chloroquine and cultured overnight. The following day the cells were fed 10uM EdU under regular growth conditions for one hour and then fixed and permeabilized. EdU was used to was visualize proliferating cells using The Click-iT® EdU Alexa Fluor® 488 Imaging kit (pink). Cells were counter stained with 0.5ug/mL anti-LC3B with a goat anti rabbit Alexa Fluor® 647 secondary (Green), mouse anti alpha tubulin with a goat anti mouse Alexa Fluor® 555 secondary (Cyan) and 1ug/mL Hoechst 33342 (Blue). Cells were imaged on a Molecular Devices ImageXpress High content imager