(A) Expression of p53 protein in human CLL cells at 1, 4, and 24 hours after incubation with medium or 0.18 μmol/L of flavopiridol. The cells were obtained from CLL patients after obtaining informed consent, isolated, and cultured at 5 × 106/mL in medium or 0.18 μmol/L of flavopiridol. Cell lysates were prepared and protein concentration was quantified using the BCA method (Pierce). Twenty-five micrograms of protein/lane from the CLL cell lysates was loaded onto a 10% SDS-PAGE gel and electrophoresed. The p53 protein (identified by an arrow) was detected using an anti-p53 monoclonal antibody (Oncogene). (B) To assess for equal lane loading, gel 6A was reprobed with antiactin polyclonal antibody (Santa Cruz), as depicted. Equal lane loading was also confirmed using Fast Green Staining (not shown).
