Fig. 2.

Mutant p53 signaling via ENTPD5 is required for FN-mediated cell adhesion, migration, and invasion. a Adhesion kinetics of MIA PaCa-2 cells to fibronectin (FN) following siRNA-mediated depletion of p53, ENTPD5, ITGA5 or ITGB1. Adhesion is expressed as the percentage of the seeded cell number. b Transwell migration and invasion of MIA PaCa-2 cells depleted of p53, ENTPD5, or ITGA5, assayed in the presence or absence of FN. Numbers of migrated and invaded cells were normalized to seeding controls and expressed as the percentage of mock-treated cells. c Adhesion kinetics of MIA PaCa-2 pIND-ENTPD5 cells (with tet-inducible expression of ENTPD5) to FN. Cells were induced with doxycycline (+ tet), transfected with siRNAs as indicated and analyzed as in a. d-e Transwell migration and invasion of MIA PaCa-2 pIND-ENTPD5 cells in the presence or absence of FN. Cells were induced with doxycycline (+ tet) as indicated, transfected with siRNAs, and analyzed as in b. d Quantification of assay results. e Representative images of crystal violet-stained transwell inserts containing FN. Proliferation control wells document comparable cell seeding and viability. All graphs show the mean ± SD of n = 3 independent experiments. Significance was tested by two-way ANOVA followed by Dunnett’s multiple comparisons test: *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ns, not significant