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The Tudor-SN-shRNA stable cells, the Scr-cont cells or the parental MDA-MB-231 cells were cultured to confluence and scratched in six-well plates. As shown in Fig. 2A, the parental and Scr-cont MDA-MB-231 cells migrated more rapidly into the wounded area, while knockdown of endogenous Tudor-SN protein (Tudor-SN-shRNA) in the cells resulted in the retardation of the wound healing. The results were quantified by image J software and demonstrated in Fig.2B. Statistical analysis demonstrated that compared with Scr-cont and parental MDA-MB-231 cells, the migration distance of Tudor-SN-shRNA MDA-MB-231 cells reduced by 71.4% and http://www.selleckchem.com/products/SRT1720.html 67.0% at 24 hr, and 66.7% and 68.7% at 48 hr, respectively (P? http://www.selleckchem.com/products/Adrucil(Fluorouracil).html examined by in vitro invasion assay. The number of cells that invaded the Matrigel layer and attached to the lower side of the filter was counted. As shown in Fig. 2C and D, knockdown of Tudor-SN in MDA-MB-231 (Tudor-SN-shRNA) cells led to an average of 68% and 62% decreased invasiveness, compared with Scr-cont or parental 231 cells, respectively (P? http://en.wikipedia.org/wiki/MERTK in HeLa cells as presented in Table 1, and miR-127 was one of the downregulated miRNAs by Tudor-SN. To further validate the results of microarray in breast cancer cells, we examined the effects of Tudor-SN on miR-127 expression on the basis of introducing exogenous miR-127 into the MDA-MB-231 cells. The pre-miR127 plasmid (GFP-pre-miR-127) was first transfected into MDA-MB-231 cells. After 36 hr incubation, the cells were divided into four different plates and then transfected with pSG5-Tudor-SN-Flag, Tudor-SN siRNA, scramble control (Scr) or pSG5 plasmid as control, respectively. Total cell lysate or RNA was collected 48 hr post-transfection and analyzed from different transfected cells. As shown in Fig. 3A, the protein level of endogenous ��-actin from different lysates were comparable, the ectopically expressed Tudor-SN was detected by blotting with anti-Tudor-SN antibody (upper panel, Lane 2). On the other hand, the endogenous Tudor-SN was efficiently depleted (upper panel, Lane 4) by siRNA.