Figure 6
(A) The left and center histograms report the results of the proliferation assays performed on MM cells (U266) stained with Carboxyfluorescein succinimidyl ester and co-cultured with NC-DCs or 29b-DCs, or cultured in the presence of conditioned medium obtained from MM/NC-DCs and MM/29b-DCs co-cultures, respectively. The right histogram represents the number of MM cells attracted by the conditioned medium obtained from MM/NC-DCs and MM/29b-DCs co-cultures (migration assay). The blots on the right side of the panel represent the main survival and proliferation signaling (ERK, AKT, SRC and p21) in MM cells evaluated after 48 h co-culture with NC-DCs or 29b-DCs by western blotting. *P<0.05. (B) Left, evaluation of DNA damage response activation in MM cells (U266) co-cultured with either NC-DCs or 29b-DCs in western blotting; right, evaluation of g-H2AX nuclear foci (DNA double-strand break markers) in confocal microscopy in MM cells (U266) co-cultured with either NC-DCs or 29b-DCs. (C) This cartoon shows the main molecular and functional changes induced by miR-29b enforced expression in DCs in the context of MM microenvironment. (a) The ‘normal’ pathologic status, in which MM cells induce a downregulation of miR-29b in DCs thus promoting an inflammatory microenvironment that leads to a survival advantage. (b) The changes that we demonstrated occuring after miR-29b transfection in DCs.
