Fig. 4. N-ethylmaleimide (NEM) completely blocks the formation of MDC-labeled vacuoles. Cells were pre-incubated for 15 minutes in the presence or the absence of 50 µM NEM. Autophagy was subsequently induced by incubating the cells in starvation medium for 60 minutes. Cells were labeled with 0.05 mM MDC for 10 minutes at 37°C and then washed four times with cold PBS pH 7.4. Cells were immediately analyzed by fluorescence microscopy as indicated in Fig. 1.

Return to article