Fig. 4
A p190A reduces invadopodia formation and gelatin degradation in BC cells. (A) Representative immunofluorescence (IF) images and quantification of invadopodia formation in bladder cancer cells. Invadopodia were identified by co-localization of F-actin and cortactin (white arrows). Scale bar, 10 μm. To quantify invadopodia formation, F-actin and cortactin double-positive puncta were counted from 8–10 random microscopic fields per condition. The number of invadopodia per cell was calculated by dividing the total number of invadopodia by the total number of cells. (B) Representative images and quantification of gelatin degradation in bladder cancer cells. Assessment of the functional consequences of altered invadopodia formation was investigated by fluorescent gelatin degradation assays. Areas of matrix degradation appear as dark regions beneath or within cell clusters (white arrows). Scale bar, 50 μm. Degradation was quantified from 8–10 random fields per condition. Gelatin degradation capacity of the cells was quantified by measuring the degradation area per cell and the degradation area per cell was calculated. N-numbers indicate total cell number from three independent experiments (RT4-shScr: n = 949; RT4-shp190A-1: n = 1185; RT4-shp190A-2: n = 1722; RT4-shp190A-3: n = 839; BFTC-EV: n = 702; BFTC-p190A: n = 1218; T24-EV: n = 1059 and T24- p190A: n = 934). A, B All experiments were performed at least in triplicates (n = 3). Quantification was based on the results of three independent experiments, and results are presented as mean ± SEM. ANOVA (RT4) and Student’s t-test (BFTC, T24) were used for the statistical analyses. (* p < 0.05, ** p < 0.01).
