Figure 5

Expression levels of genes involved in cell cycle suppression ( KLF4 , P21^CIP), cell proliferation ( MYC, CCND1, BCL3, SP1 ) and apoptosis ( AATF ) in pediatric acute lymphoblastic leukemias. (A-B) qRT-PCR analysis for expression level of genes including P21^CIP, BCL3, MYC and AATF in B- and T-cells from patients with pediatric ALL compared with the corresponding controls. Expression was normalized to β-actin and each bar represents mean ± S.D of the experiment performed in triplicate; **P < 0.01; *P < 0.05 relative to control B and T cells. (C-E) Western blotting to determine protein expression levels of P21^CIP(C), CCND1 (D) and SP1 (E) in both B- and T-lineage blasts compared with corresponding controls. Expression was normalized to β-actin and protein band intensities were determined using Scion Image Analysis Software. Each bar represents mean ± S.D. of the experiment performed in triplicate; **P < 0.01; *P < 0.05 relative to control B and T cells. (F) SP1 transcription factor promoter activity in both B-ALL and T-ALL samples compared with the corresponding controls. B- and T-lymphoblasts were transfected with reporter plasmids containing SP1 response elements, incubated for 72 h at 37°C in humidified 5% CO₂ atmosphere. The experiments were repeated thrice and results were reported as relative β-gal activity. Each bar represents mean ± S.D of the experiment performed in triplicate *P < 0.05 relative to control.