Fig. 4

Knockout of AGR2 alters the cell-cell and cell-matrix adhesion and increases the invasiveness of A549 cells. a Control A549 cells show epithelial-like colonies with high level of cell-cell adhesion; both AGR2 knockout clones show reduced cell-cell adhesion and have spindle-shape morphology. Phase-contrast microscopic images were taken at day 5 after cell seeding. Single cell suspensions were placed on b uncoated or c Matrigel-coated tissue culture plates for 2 h for adhesion assay and for 90 min for cell-matrix adhesion assay. The error bars represent the standard deviation of the mean determined from three independent experiments, each performed in duplicate. d Cells were plated in the upper chamber of a CIM-16 plate coated with Matrigel. Real-time invasion of cells was measured on an Xcelligence Real-Time Cell Analyzer DP analyzer. Representative graph (left) for A549 and both clones of A549 KOAGR2 (G2, G9) shows invasiveness of A549 cells in relation to AGR2 expression. Each clone was determined in triplicate. Resulting graph (right) obtained from two independent experiments (representative experiment is shown on the left) shows cellular invasiveness determined at 60 h. Data are presented as the mean +/− standard deviation (n = 5). Statistical significance (P < 0.05) is indicated by asterisks