Figure 3. Silencing of hPEBP4 sensitized the primary B-ALL cells to R-CDC effect ex vivo.
A. 5×106 Primary B-ALL cells from 8 different patients were transfected with hPEBP4 siRNA or scrabble siRNA using the Amaxa nucleofector II (Amaxa Biosystems, Koln, Germany) as recommended by the manufacturer. 24 hours after transfection, Real-time PCR data confirmed the downregulation of hPEBP4. B. Expression of hPEBP4 was determined in B-ALL cells by flow cytometry after intracellular staining. Representative flow cytometry from patient 6, gated on CD20+ cells. the purity of the malignant cells were measured by CD20 expression. C-D. R-CDC induced more death in hPEBP4-silent B-ALL cells compared with control (P = 0.0098). 24 hours after transfection, the transfected B-ALL cells were preincubated with 20 µg/mL rituximab for 60 min, and then stimulated by 2% NHS for 1 hr, followed by PI staining.