Figure 1

Effect of PRL-3 siRNA on PRL-3 expression. (A) The secondary hairpin structure of pRNA-U6.1/Neo-PRL-3 siRNA transcripts. (B) Photographs of transfection efficiency of B16-BL6 cells. B16-BL6 cells were transfected with pEGFP-N1 for 24 h and then photographed under fluorescent microscopy (right) and light microscopy (left). (C) Expression of Myc-PRL-3 protein was examined by Western blot in 293T cells cotransfected with Myc-PRL3 expression vector (pTARGET) and Luc siRNA, PRL-3 siRNA-1, −2, −3, or −4 RNAi vector (pRNA-U6.1/Neo). (D) Effect of PRL-3 siRNA on endogenous PRL-1, PRL-2, and PRL-3 expression in B16-BL6 melanoma cells. B16-BL6 cells were untreated or transfected with Luc siRNA and PRL-3 siRNA-1 (1 and 2 µg). After treatment at 37°C for 24 h, PRL-1, PRL-2, and PRL-3 mRNAs were studied by RT-PCR; levels of GAPDH expression are shown as an internal control. (E) Real-time quantitative PCR evaluation of PRL-3 siRNA’s effect on endogenous PRL-3 expression in B16-BL6 melanoma cells. Data are mean ± SEM of three independent experiments, and each experiment includes triplicate sets. **P < 0.01 vs. Luc siRNA control.