Figure 5

Expression of HOXA10 reversed cell invasive property caused by miR-135a in BT549 cells. (A) Invasion assay of MDA-MB-231 cells 48 h after cotransfecting pS-135a or pS-negative and pcDNA HOXA10 contains a full-length HOXA10 cDNA, or pcDNA HOXA10-mu-3'UTR, mutated in the miR-135a binding site in the 3'-UTR. Images of invasive MDA-MB-231 cells (top panel). (B) Invasion assay of BT549 cells 48 h after cotransfecting 135a inhibitor or inhibitor N.C and HOXA10 siRNA (siHOXA10) or Control siRNA (siControl). Images of invasive BT549 cells (top panel). (C) Western blot of HOXA10 protein in BT549 cells 48 h after expressing HOXA10 (full-length or 3'-UTR region deleted) by transfection. Anti-GAPDH antibody was used as a loading control (bottom panel). (D) Invasion assay of BT549 cells 48 h after transfection. pcDNA NC was used as negative control, pcDNA HOXA10 contains a full-length HOXA10 cDNA including its 3' UTR, and pcDNA HOXA10-de-3'UTR contains the HOXA10 coding sequence with the 3'-UTR region deleted. Images of invasive BT549 cells (top panel). n = 3, *, p < 0.05, significantly decreased migration or invasion.