Figure 4.

Effects of FAM188B knockdown on several metastatic characteristics in lung cancer cell lines. (A–E) Cells were transiently transfected with either si-NC or si-FAM188B for 24 h, followed by invasion assay (A), migration assay (B), adhesion assay (C), wound healing assay (D), and 3D culture (E) as described in the “Materials and Methods” and representative images are shown. For relative invasion and migration analysis, images were scanned to quantify three different areas (right panels in A,B). For adhesion assay, 5 × 10³ cells were plated in 96-well plate, and at indicated times, floating cells were removed by washing, and attached cells were measured by MTS assay (C). For wound healing assay, confluent cells were grown in 12-well plate, and cell monolayers were scratched with a pipet tip, and then cells were incubated for 24 h (D). For 3D culture, 5 × 10³ cells in 400 μL of 5% Matrigel were plated in the Matrigel-coated 8-well chamber as described in the “Materials and Methods”. Colony images were captured on indicated days (E). Scale bars = 100 μm. Error bars represent standard deviations of the mean of three measurements (* p < 0.05, ** p < 0.01, versus si-NC). These experiments were performed three times independently with similar results.