Fig. 8.

Monitoring of CYTO-ID dye fluorescence signal in autophagy compartments in IMR-32 cells. Localization of the CYTO-ID and Hoechst 33342 fluorescence dyes in IMR-32 cells was assessed using fluorescence microscope. IMR-32 cells were pre-treated with 10 μM CQ for 1.5 h and subsequently treated with 0.1 μM MK-5108 or DMSO for 48 h. The cells were then stained with CYTO-ID and Hoechst 33342, fixed, and visualized under fluorescence microscope. Relative mean fluorescence intensity of CYTO-ID-stained autophagic compartments was quantified in five randomly selected photomicrographs (taken using a ×40 objective). Scale bar 25 μm. C—control, DMSO-treated cells, MK—MK-5108-treated IMR-32 cells, CQ—chloroquine-treated IMR-32 cells, PC—positive control, IMR-32 cells grown in amino acids-free medium for 24 h