CYRI proteins affect the invasion ability of cancer cells through integrins. (A and B) Organotypic assay of control pLKO versus CYRI DBKO A-673 cells stained with hematoxylin and eosin. (B) Quantification of the invasion index (the area invaded by cells divided by the total area of the plug). Data from at least three independent experiments. Each experiment is color-coded. Mean ± SD. Statistical analysis using ANOVA with Tukey’s multiple comparison test. (C and D) Inverted invasion assay comparing the invasion capacity between the control pLKO and the CYRI DBKO A-673 cells in the presence or absence of the integrin α5β1-blocking antibody IIA1. Scale bar = 100 µm. (D) The graph shows the quantification of the invasion index (the area covered by cells beyond 10 µm into the plug). Data from at least three independent experiments. Each experiment is color-coded. Mean ± SD. Statistical analysis using two-tailed unpaired t test. (E and F) Phase-contrast images of the soft agar assay comparing the capacity for anchorage-independent growth of the control pLKO and the CYRI DBKO A-673 cells. The size of the spheroids is quantified in F. Scale bar = 5 cm. Data from three independent experiments. Each experiment is color-coded. Mean ± SD. Statistical analysis using ANOVA with Tukey’s multiple comparison test. ns, P > 0.05.