Figure 5.

Depletion of uL3 enhances autophagic flux in HCT 116^p53−/− cells and the restoration of uL3 in uL3ΔHCT 116^p53−/− cells rescues this effect. Representative z-stack images of uL3ΔHCT 116^p53−/− treated with 25 µM CQ or 1 µM RAPA for 4 h or transiently transfected with 1 µg of pHA-uL3 for 24 h, alone or in combination. Cells were fixed and double stained with LC3B and LAMP1 antibodies. Nuclei were counterstained with Hoechst. Single-color fluorescence images of LC3B positive autophagosomes (green) and LAMP1 positive endosomes and/or lysosomes (red) are presented in the 1st and 2nd columns, respectively, and a merged view of these 2 proteins is shown in the 3rd column. Higher magnification views of the boxed area from the merged images are shown. Scal bar, 10 μm.