Fig. 2. VAL potently promotes invasion and metastasis in LAD.

a Relative luciferase activities of vector-control or VAL-overexpressing LAD cells adhering to the Matrigel. b Representative images and quantification of invading cells in five random fields of Matrigel-coated transwell assay. c The sub-G1 DNA contents of detached cells for vector-control or VAL-overexpressing A549 or H2009 cells are shown in the anoikis assay. d–f The effect of silencing VAL on cell adhesion, invasion, and suspension-induced anoikis in H1975 cells. g Representative images of H&E staining display a clear boundary and irregular invasive front between the dermal tissue and tumor tissue in the subcutaneous tumors (n = 5 per group), respectively, xenografted with vector-control and VAL-overexpressing LAD cells. Black arrows mark the direction of tumor invasion with invasive front marked by dashed line. h LAD cells overexpressing VAL or silenced with VAL or vector-control cells labeled with luciferase expression were injected via cardiac ventricle into nude mice (n = 5 per group). Representative bioluminescent images of systemic metastasis are shown. i Frequencies of formation of metastatic lesions in nude mice (n = 5 per group) intracardially injected with different cell numbers of vector-controls, VAL-overexpressing, or VAL-silenced LAD cells. j Representative images of metastatic bone lesions in mice (n = 5 per group) intracardially injected with the indicated cells were confirmed by micro-CT imaging and H&E staining. Scale bar: 100 μm (b, e, g, j). Scatter dot plots represent the means ± SD derived from three independent experiments. Statistical analyses were performed by two-tailed unpaired Student’s t test (a–f). Source data are provided as a Source Data file.