FIGURE 5.

NEGR1 and UCA1 are necessary for TGFβ-induced tumorigenic events. A, LM2-4 cells were transiently transfected with control siRNA (siCTL) or siRNA targeting NEGR1 (siNEGR1) or UCA1 (siUCA1) and treated with TGFβ. Monolayers were wounded and analyzed for cell migration. Representative images of three independent experiments are shown. B, LM2-4 cells transfected with siCTL, siNEGR1, or siUCA1 were plated on transwell filters to assess cell migration. Migrated cells are shown as the average number in 10 random fields over two independent experiments + S.E. C, LM2-4 cells were transfected with siCTL, siNEGR1, or siUCA1 and then grown under anchorage-independent conditions in the presence of TGFβ to examine primary mammosphere formation. Representative images are shown, and three independent experiments from each condition were quantitated, measuring the number of colonies formed and the size of each colony. Black error bars represent the average + S.E., and red error bars represent the average ± S.E., **, p < 0.005; ***, p < 0.0001 (t test).