Figure 9.

UNC119A silencing malignant transformation in NIH3T3 cells expressing KRASB G12V and human lung cancer A549 cells. A, control and Myc-KRASB G12V-expressing NIH3T3 cells were transfected with control siRNA (siCont) or UNC119A siRNA (siUNC119A#1). Soft-agar colony formation assay was performed. Colony area was evaluated in 15 independent fields. Dot plots exhibit each value. Horizontal bars show the averages. ***, p < 0.001. The immunoblottings demonstrate the expression of Myc-KRASB G12V and the suppression of UNC119A. B, C, and D, A549 cells were transfected with control siRNA (siCont), UNC119A siRNA (siUNC119A#1), and KRAS siRNA (siKRAS). Soft-agar colony formation, sphere formation, and transwell (migration and invasion) assays were performed. The immunoblottings demonstrate the suppression of UNC119A and KRAS. B, colony formation was evaluated as described for panel A. C, cell aggregates larger than 50 µm were defined as spheres, and the number of spheres was counted in 5 independent fields. D, cells were cultured on transwell inserts without or with Matrigel. 36 h later, the membranes were fixed and the cells on the lower surface were stained with crystal violet. The intensity of staining was quantified by ImageJ in 5 independent fields. ***, p < 0.001. Scale bars, 100 μm (A, B, and C) and 200 µm (D). Three experiments were performed. The data are means with S.E. Numbers of samples are shown. Statistical analyses were performed with one-way ANOVA with Tukey's test.