Expression profiling MOLT-4 treated with MABL

Analysis of MOLT-4 cells at various time points up to 6 hours following treatment with mouse anti-CD47 antibody (MABL) and goat anti-mouse IgG (GAM) as the crosslinker of MABL. MABL induces apoptosis in CD47-positive MOLT-4 cells. Cell death signals via CD47 ligation were analyzed by using Affymetrix Human Genome U133A microarray. MOLT-4 cell line was obtained from the American Type Culture Collection (ATCC) and grown in RPMI1640 medium (SIGMA) with 10% FBS(JBS). It was maintained at 37 degree C and 5% CO2. Cells were washed with PBS(-) followed by treatment with 10 ug/mL MABL (anti-CD47 antibody) for 0, 0.5, 1, 3 and 6 h. Total RNA was isolated and converted to cDNA using a Superscript Double Stranded cDNA synthesis kit (Invitrogen). Fragmented cDNA was hybridized to Affymetrix Human Genome U133A array and gene expression signals were analyzed. Probes showing >2 or <0.5 fold-change for both replicates compared with the control (geometric mean of 0h replicate signals) were selected at any time.