Resveratrol causes accumulation of MM cells in sub-G1 phase, increases release of Bax protein accumulation, and activates caspase-3. (A) U266 cells (2 × 106/mL) were synchronized by incubation overnight in the absence of serum and then treated with 50 μM resveratrol for 0, 12, or 24 hours, after which the cells were washed, fixed, stained with PI, and analyzed for DNA content by flow cytometry. Results typical of 3 independent experiments are shown. *P < .05. (B) MM1.S cells (2 × 106/mL) were synchronized by incubation overnight in the absence of serum and then treated with 50 μM resveratrol for 0, 12, or 24 hours, after which the cells were washed, fixed, stained with PI, and analyzed for DNA content by flow cytometry. Results typical of 3 independent experiments are shown. *P < .05. (C) U266 cells (2 × 106/mL) were treated with 50 μM resveratrol for the indicated times, and whole-cell extracts were prepared, separated on sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), and subjected to Western blot using antibody against Bax. The same blots were stripped and reprobed with β-actin antibody to show equal protein loading. (D) U266 cells were treated as described in panel C, and whole-cell extracts were prepared, separated on SDS-PAGE, and subjected to Western blot using antibodies against indicated proteins. The same blots were stripped and reprobed with β-actin antibody to show equal protein loading. (E) MM1.S cells were treated as described in panel C, and whole-cell extracts were prepared, separated on SDS-PAGE, and subjected to Western blot using antibody against PARP. The same blots were stripped and reprobed with β-actin antibody to show equal protein loading. The results shown are representative of 3 independent experiments.
