MVs modulate GSK3β/β-catenin signaling in BMSCs. (A) The BMSC lysates stimulated with MVs described in Figure 4A were analyzed for AKT-mediated phosphorylation of GSK3β and β-catenin at Ser552 by Western blot by the use of specific antibodies. Phosphorylation of AKT is also shown for comparison. (B) Translocation of β-catenin was visualized in BMSCs stimulated with MVs from CLL patients (MV1 and MV2) by the use of a specific antibody to β-catenin and nuclear stain, DAPI under confocal microscope (magnification level is indicated by the horizontal bar). (C) AKT-mediated phosphorylation of β-catenin at Ser552 and GSK3β was further examined in BMSCs stimulated with MVs for 24 or 48 hours by Western blotting. Expression of cyclin D1 and c-myc, downstream targets of GSK3β/β-catenin, was shown in BMSCs after stimulation with MVs (MV22 and MV23) compared with the unstimulated controls in Western blot analysis.