Fig. 6.

Quantitative analysis in pulse, and BFA experiments. In pulse chase experiments (A, B) protoplasts were incubated for 15 min (T₀) with 30 nmol of nanogold and then particles were removed from the medium. Samples were fixed after 15 min and 45 min. (A) The percentage of nangold internalized in different membraneous compartments were calculated as described for the time-course experiments. Pulse experiments allowed the distribution of internalized nanogold within the cell compartments to be followed until 45 min, suggesting that recycling could be a prominent pathway with respect to degradation during nanogold endocytosis. In Golgi bodies and ER much less probe was observed (B) and the percentage of labelled Golgi bodies decreases with respect to the time-course. (C) Quantitative analysis showed that in BFA-treated tobacco protoplasts incubated with positively charged nanogold, BFA affected the early phases of the internalization process. Since BFA was dissolved in DMSO, BFA effects on nanogold internalization were compared with DMSO. Standard error (SE) is shown for each experiments.