(A) Expression of bax protein in human CLL cells at 4, 24, and 48 hours after incubation with medium or 0.15 or 0.015 μmol/L of depsipeptide. The cells were obtained from CLL patients after informed consent was obtained, were isolated, and were cultured at 5 × 106/mL in medium or depsipeptide. Cell lysates were prepared and protein concentration was quantified using the BCA method (Pierce). Two micrograms of protein per lane from the CLL cell lysates was loaded onto a 10% SDS-PAGE gel and electrophoresed. Bcl-2 protein was detected using an anti–bcl-2 monoclonal antibody (Dako). Lane equivalent loading was verified by assessment with Fast Green staining (not shown). (B) Expression of total protein on nitrocellulose membrane for gel (A) after staining with Fast Green. This demonstrates equivalent lane loading for each of the time points outlined in (A).
