Figure 4

Functional consequences of silencing PRL-3 in CML cells. (A) The mRNA expressions of PRL-3 and β-actin were analyzed RT-PCR in K562-shRNA-Scramble (shC) and K562-shRNA-PRL-3 (shP) transduced cells (i). P.D. stands for primer dimer (i). Cell proliferation curves were constructed by the total viable cells in ShC- and ShP-K562 cells cultured over 8 days. Cell proliferation and viability were determined by trypan blue counting in every other day. Results represent the mean ± SD of triplicates (ii). Colony Forming Unit (CFU) assay of in ShC- and ShP-K562 cells. The experiments were duplicated. The upper panel of images was taken by a Canon EOS40D camera and the lower panel of pictures was captured in 4 × 10 magnification field under an invert microscopy. The experiments were duplicated and representative pictures were presented (iii). (iv) Comparison of tumor formation capacity in mouse xenograft models between K562-shC and K562-shP cells. (v) Average tumor weight of three K562-shC tumors. Bar indicates standard deviation (SD). (B) P210 WT and T315I cells were nucleofected using the cell line solution V (Lonza), program X-001 and nontargeting (NC) or mouse PRL-3 siRNA (Santa Cruz Biotechnology, Inc.). After 24 h, cells were subjected to conventional RT-PCR and qRT-PCR analysis of PRL-3 expression or MTS assays. Cell proliferation assessments in untreated, Imatinib-treated (5, 10 μM) P210 T315I cells transfected with either nontargeting (NC) or mouse PRL-3 siRNA for 48 h and determined by MTS assay. Results were presented the mean ± SD of 3 independent experiments. Significant value *p < 0.05 and **p < 0.01.