Figure 4From: RIZ1 is potential CML tumor suppressor that is down-regulated during disease progressionEffect of RIZ1 expression on differentiation in CML myeloid blast crisis model cell lines. (a) Benzidine staining assays comparing erythroid differentiation in K562 cells transfected with shRNA non-silencing control plasmid (K562), K562+RIZ1 cells transfected with shRNA non-silencing control plasmid (K562+RIZ1), and K562+RIZ1 cells transfected with pRIZ1shRNA (K562+RIZ1+shRNA). (b) Western analysis of RIZ1 expression in K562 transfected with shRNA non-silencing control plasmid (K562), K562+RIZ1 cells transfected with shRNA non-silencing control plasmid (K562+RIZ1), and K562+RIZ1 cells transfected with pRIZ1shRNA (K562+RIZ1+shRNA). (c) RT-PCR analysis of RIZ1 mRNA expression in ERY-1 and YN-1 transfected with shRNA non-silencing control plasmid (Con shRNA) or with pRIZ1shRNA (RIZ1 shRNA). Total RNA was reverse transcribed and cDNA amplified with RIZ1 and β-actin-specific primers. M represent DNA ladder and H2O represents RT-PCR reaction without template DNA. (d) Erythroid differentiation assay comparing ERY-1 and YN-1 after transfection with pRIZ1shRNA or shRNA non-silencing control plasmid (Con). Cell lines were transfected with pRIZ1shRNA or shRNA non-silencing control plasmid and cultured for three days. Histograms show the percentage of benzidine-positive cells that were scored by light microscopy. Error bars represent the standard deviation from three independent experiments. (e) CD33 and CD117 expression in JURL-MK1 cells as compared with JURL-MK1 cells expressing RIZ1 (JURL-MK1+pRIZ). JURL-MK1 was transfected with pRIZ1 or pcDNA3 control plasmid (con) and cultured for three days. Panel (i) shows the fluorescence intensity of phycoerythrin (PE)-conjugated antibody against CD33. Panel (ii) shows the fluorescence intensity of (PE)-conjugated antibody against CD117. Panels (iii) and (iv) show immunocytochemical staining using an anti-CD117 antibody in JURL-MK and JURL-MK1+pRIZ1 cells, respectively.Back to article page