Figure 3.

Specc1l^cGT/ΔC510 primary MEPM cells exhibit migration defects in wound-repair assays. (a–f) Phase contrast micrographs of a representative wound-repair assay performed with wild-type (WT) and Specc1l-mutant MEPM cells. While WT cells (a–c) show complete wound-closure by 36 h (h), the mutant cells (d–f) require more time. The wound area is indicated by two-headed arrows. (g) Automated confluence analysis of wound-repair experiments quantified the average percentage of wound-closure over time. Orange and blue colors indicate data obtained from WT and mutant cultures, respectively. By 24 h (dashed vertical line), WT MEPM cells close more than 66% of the wound. Solid lines represent the average of 29 independent microscopic fields, recorded in 5 independent experiments. Shaded areas indicate SEM. (h) Average cell motility speed, calculated by PIV analysis of the 29 wound-closure recordings analyzed in (g). For each time point velocity magnitudes within the entire cell-covered area were averaged. Despite approaching wound-closure after 24 h (dashed line), cells remained motile in both WT and mutant MEPM cultures. (i) Statistical analysis of data presented in (g,h) showed that the motility of mutant MEPM cells is significantly reduced in comparison to WT MEPM cells. Left: average speed of wound front, calculated within the 1–15 h time interval (p < 6.7 × 10⁻⁶, Welch’s t-test). Right: average speed of cell movements within the entire recorded time period of 40 h (p < 2.2 × 10⁻⁵, Welch’s t-test). Error bars represent SEM.