Effect of MVs from healthy and diabetic individuals on differentiation of K562 cells. (A-D) Cells were treated for 72 hours with medium (control, gray) or in vivo circulating MVs from healthy (2 μg/mL; A-B) or diabetic individuals (5 μg/mL; C-D) respectively, in the absence (blue trace) or presence of 30 μM cyclopamine (red trace). (E-H) Cells were treated for 72 hours with medium (gray) or 5 μg/mL MVs generated from PHA/PMA/act D-treated lymphocytes from healthy (E-F) or diabetic individuals (G-H) in the absence (blue trace) or presence of 30 μM cyclopamine (red trace). Detection of α_IIbβ₃ and CD42b expression was performed by flow cytometry after labeling by the corresponding antibody. Fluorescence (FL2, red fluorescence channel intensity) is expressed in arbitrary units (au). Data are representative of 4 experiments yielding similar results. (I-J) Histograms representing the increase of mean fluorescence intensity (± SEM) reflecting the expression level of α_IIbβ₃ (I) and CD42b (J), in control cells, or in cells treated with circulating MVs from healthy (2 μg/mL, white bars) or diabetic individuals (5 μg/mL, black bars) or 5 μg/mL MVs generated from PHA/PMA/act D–treated lymphocytes from healthy (hatched bars) or diabetic individuals (dotted bars) in the absence or presence of 30 μM cyclopamine (cycl). *P < .05, **P < .01, ***P < .001, significantly different from nontreated control cells; †P < .05, ††P < .01, significantly different from MV-treated cells.