Figure 5.

c‐Src/c‐Yes/Fer are crucial for microRNA (miR)‐129‐1‐3p‐mediated regulation of tumor progression. A, HCT116 cells transfected with the c‐Src and c‐Yes ORFs and a Fer mutant resistant to miR‐129‐1‐3p cDNA were treated with control (cont)‐miR or miR‐129‐1‐3p, and total cell lysates were immunoblotted with the indicated abs. The relative expression levels of each protein are shown below the panels. B, Soft‐agar colony formation assays of the cells in (A). Relative colony numbers ± SD were obtained from 3 independent experiments. C, HCT116 cells in (A) were subjected to immunocytochemistry. F‐actin (red), vinculin, a marker of focal contact (green), and DAPI (blue) were analyzed by immunostaining HCT116 cells grown on fibronectin‐coated dishes. Scale bar = 20 µm. D, HCT116 cells used in (A) were subjected to adhesion assay on fibronectin‐coated dishes. E, In vitro invasiveness of the HCT116 cells in (A). Cells were seeded in Matrigel invasion chambers. After 48 h, the membranes were detached, and the cells were stained and counted. Relative numbers of invading cells per mm² ± SD were obtained from 3 independent experiments (B, D, E). *P < .05, **P < .01 by Student’s t test. n.s., not significant