Fig. 5. Vinblastine-treatment causes a marked increase in the size of autophagic vacuoles. (A) CHO cells were incubated in full nutrient conditions (A,C,E) or in starvation medium (B,D,F) for 3 hours, in the presence (C,D,E,F) or absence (A,B) of 50 µM vinblastine. In E and F cells were pre-incubated for 3 hours with 10 mM 3-MA. Both starved and control cells were subsequently incubated 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. (G) The number and size of MDC-labeled vacuoles displayed in Fig. 5A-D were quantified by Methamorph software, using the integrated morphometry analysis. Data represent the mean±s.e.m.
