LACSLCs derived from A549 cells exhibit highly invasive and migratory capabilities.

<p>(<b>A</b>) Morphology of sphere-LACSLCs derived from lung cancer A549 cell line. Representative images from light microscopy (monolayer cells 10×, LACSLCs 20×) are shown. (B) Wound healing assay for cell migration of A549 LACSLCs and A549 monolayer cells. Representative images of cell migration into the wounded area at 0 and 16 hours post-injury (<i>left panel</i>) are shown. Quantitative analysis shows wound repair capability of migrating cells at 16 h post-injury (<i>right panel</i>). (<b>C</b>) Comparison of invasive capability in A549 LACSLCs and monolayer cells detected by transwell assay. Representative images of invading cells visualized by crystal violet staining (20×) (<i>left panel</i>). Quantitative analysis of cell invading capacity at 24 h after seeding (<i>right panel</i>). (<b>D</b>) Histological images by H&E staining under light microscopy. Tumors formed by A549 LACSLCs were highly invasive and the tumor cells invaded neighboring muscle layer (red arrows; 20×). All experiments were carried out at least in triplicates and the data are presented as the mean ± SEM. Student <i>t</i> test was performed to evaluate the difference.</p

Examination of POU5F1 and MMP-2 by immunostaining and Western blot analyses.

<p>(<b>A</b>) Immunostaining of POU5F1 and MMP-2 observed by confocal scanning microscopy. (<b>B</b>) Expression of stem cell transcription factor POU5F1 was detected by Western blot. (<b>C</b>) Immunohistochemical analysis of MMP-2 protein in tumor tissues formed by A549 LACSLCs and monolayer cells. Representative images of MMP-2 expression by IHC staining (20×) (<i>left panel</i>). Quantitative analysis of MMP-2 protein levels in tumor tissues formed by A549 LACSLCs and monolayer cells. All experiments were carried out at least in triplicates and the data are presented as the mean ± SEM. Student <i>t</i> test was performed to evaluate the difference.</p

The expression of POU5F1 and MMP-2 in 55 cases of lung adenocarcinoma.

<p>^a Mean age.</p