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. 2015 Apr 12;6(15):13772–13789. doi: 10.18632/oncotarget.3830

Figure 7. Exosomes released by multiple myeloma cells support osteoclast precursors survival and inhibit apoptosis.

Figure 7

A. Facs analysis of Annexin V iodide staining of Raw264.7 cells treated for 24 hours with 25ng\ml hrRANKL, 1% DMSO or 25 μg/ml of U266- and MM1s-derived exosomes B. Caspase 3/7 activity assay was performed in Raw264.7 cells untreated or treated for 24 hours with 25ng\ml of hrRANKL and with 25 μg/ml of U266- and MM1s-derived exosomes. Luminescence was measured using the Caspase 3/7 Glo assay. Results are expressed as percentage of caspase activity compared to untreated cells. Averaged values of three independent experiments are plotted including ±S.D. Sidak test: *,EXO-U266/MM1s vs Untreated (*p < 0.05); °,hrRANKL vs Untreated (°p < 0.05); C. Quantitative RT-PCR of mTRAP in Raw264.7 cells untreated or pretreated with a caspase-3 inhibitor (50μM Z-DEVD-FMK) 1h before and during hrRANKL or MM-exosomes treatment. Values are expressed as fold of control and are the mean of three different experiments. Sidak test: *,EXO-U266/MM1s + ZDEVD vs EXO-U266/MM1s (*p < 0.05); °,hrRANKL+ZDEVD vs hrRANKL (°p < 0.05); + D. Western blotting analysis of p-AKT, TOT-AKT, Bcl-xl and Survivin in Raw264.7 cells untreated or treated with 25ng\ml of hrRANKL (positive control) or with 25 μg/ml of U266 or MM1s exosomes (Exo U266/MM1s). β-actin was used as loading control.