Figure 3

Effect of silencing BCR-ABL fusion gene and STAT3 on PRL-3 expression. (A) Quantification of BCR-ABL and PRL-3 mRNA by qRT-PCR in K562 cells transfected with b3a2_1 siRNA and non-targeting control (NC) siRNA. Two million cells were nucleofected with 30 nM b3a2_1 siRNA or NC siRNA together with pmaxGFP (Lonza, Switzerland) as an indicator of transfection efficacy. The solution V and program T-016 were used as recommended by the manufacturer (Lonza). The transfection efficacy was about 80%. RNAs were extracted 24 h after transfection. Primer sequences, siRNA sequences and qRT-PCR methods were described as Scherr et al.[24] and Zhou et al.[19]. (B) qRT-PCR quantification of STAT3 and PRL-3 gene expression in K562 cells transfected with STAT3 siRNA (Santa Cruz Biotechnologies, Inc., CA, USA) and NC siRNA in a Nucleofection device as described in (A). The primer sequences of STAT3 are as following: 5’-AGGATGGCCCAATGGAATCAGCTA-3’ (sense) and 5’-AGCGGCTATA CTGCTGGTCAATCT-3 (antisense).