Fig. 7. MCL exosomes internalized by cholesterol dependent endocytosis pathway.
(A) Jeko-1 cells were pre-treated with 10μM Gefitinib, 80μM Dynasore, or 40μg/ml Nystatin for 3h 30 min and 60 min, respectively. For cholesterol synthesis Jeko-1 cells were cultured in serum free RPMI media supplemented with 0.1% BSA and 20mM Hepes in the presence of 2uM Simvastatin for 16 h. PKH-67 labeled Jeko-1 exosomes were incubated with untreated (control) or treated cells for 60 min and internalization was measured by flow cytometry analysis. The data are expressed as the percent of cells that uptake labeled exosomes as mean ± SD of three independent experiments. (B) PKH-26 labeled Jeko-1 exosomes were incubated with untreated (control) or treated cells (Dynasore or Nystatin) and internalization was measured by confocal microscopy. Jeko-1 cell membrane was labeled with AlexaFluor 488-anti CD44. Scale bar is 25μm.