The following antibodies were used in our experiments: mouse anti-Hsp70 (Enzo Life Science), rabbit anti-NfkB (Santa-Cruz), mouse anti-CD63 (Millipore), mouse anti-Tsg 101 (Abcam), mouse anti-Annexin V (Santa-Cruz), and goat anti Histone H3 (Santa Cruz).
Exosome preparations from MM pool were obtained as described before. Samples were normalized for protein concentration (Bradford Assay) when possible (in alternative equal volumes of each sample were loaded on a acrylamide–bisacrylamide gel), boiled in reducing SDS sample buffer (80 mM Tris, pH 6.8, 2% SDS, 7.5% glycerol, 0.01% Bromophenol blue) supplemented with 2% 2-mercaptoethanol (Sigma) for 5 min at 95 °C and separated by SDS–PAGE on a acrylamide/bisacrylamide (10% or 12.5%) gel and analyzed by WB as previously described24 (link).
Exosome preparations from MM pool were obtained as described before. Samples were normalized for protein concentration (Bradford Assay) when possible (in alternative equal volumes of each sample were loaded on a acrylamide–bisacrylamide gel), boiled in reducing SDS sample buffer (80 mM Tris, pH 6.8, 2% SDS, 7.5% glycerol, 0.01% Bromophenol blue) supplemented with 2% 2-mercaptoethanol (Sigma) for 5 min at 95 °C and separated by SDS–PAGE on a acrylamide/bisacrylamide (10% or 12.5%) gel and analyzed by WB as previously described24 (link).