Fig. 2

PI3K/AKT signaling pathway inhibition and regulation of miR29b and DNMT3B expression in MM plasma cells (U266 and RPMI 8226 cells). Cells were cultured in the presence of 10 µg/mL anti-CD28-loaded particles, alone or with PI3K inhibitor (25 µM LY294002 48 h; 0.5 µM buparlisib 24 h). A Representative western blots of PI3K/AKT pathway components under CD28 stimulation alone or with PI3K inhibition. B Densitometric analysis of the experiment shown in (A). Values were normalized to β-actin and then to values for CD28-stimulated cells (dotted line). C Plasma cell levels of miR29b and DNMT3B after PI3K inhibition normalized to those of RNU44 or GAPDH, respectively, and expressed as fold change relative to the average value for CD28-stimulated cells. D Representative western blot and E densitometric analysis of DNMT3B expression after CD28 stimulation and PI3K inhibition. Chart data are mean and SD for 7 independent experiments. *P < .05, Wilcoxon signed-rank test