Fig. 1. CTGF expression enhances migratory ability in human breast cancer cells. (A) Expression of CTGF was detected in BT474, BT483, T47D, MDA-MB-453, MCF-7, MDA-MB-231, and MDA-MB-435 cells using northern blotting. (B) The migratory ability of the breast cancer cells was tested using a wound healing migration assay. Cells were seeded at confluence under normal culture conditions for 24 hours. Monolayers were wounded by scratching with a pipette tip. Images were taken at 20x magnification. Three wells per experiment were counted and each experiment was repeated three times, error bars are the corresponding upper 95% confidence intervals. (C) CTGF expression levels in transfected MCF-7 and MDA-MB-231 cells. Expression of CTGF was detected in MCF-7/neo, MCF-7/CTGF, MDA231/neo, and MDA231/AS cells by RT-PCR (upper panels) and western blotting (lower panels). (D) Effects of CTGF on cellular growth. Cells were seeded on 24-well dishes and cell growth was assayed using a MTT assay. (E,F) Effects of CTGF on the migratory abilities of MCF-7/neo, MCF-7/CTGF, MDA231/neo, and MDA231/AS cells. The migratory ability was measured by using a Boyden chamber assay. Each of the transfected cells was tested in three separate experiments with incubations conducted in triplicate. Columns show the means of three independent experiments, and the error bars are the corresponding upper 95% confidence intervals. Asterisks denote a statistically significant difference in migratory ability of cells transfected with sense- or antisense-CTGF compared to cells transfected with the empty vector (^*P<0.05, ^**P<0.01, two-tailed Student's t-test).