Galectin-9 depletion diminishes DC 3D migration. (A and B) WT or gal9 KD moDCs were matured and embedded within a 3D collagen matrix containing either a MEL624 or a BLM melanoma cell spheroid. The median cell velocity (A) and the MSD (B) were calculated. Data represent the mean value ± SEM of a representative donor. At least 20 cells were analyzed per condition. (C–E) Migration pattern of WT or gal9 KD moDCs either immature (at day 6 of DC differentiation), mature (at day 8), and mature in the presence of a tumor spheroid (at day 8) was analyzed for one representative donor. The median cell velocity (C), the MSD (D), and the Euclidean distance reached after 60 min of tracking (E) were assessed. In D, m = mature DCs; i = immature DCs. Graphs represent the mean value ± SEM of one representative donor. At least 25 cells were analyzed per condition. One-way ANOVA was performed to compare WT and gal9 KD moDCs. n.s., P > 0.05; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.