Figure 2: CHC17 or CLC depletion decreases HeLa and H1299 cell migration.

Wound-healing assays were performed in cells transfected with siRNA against CHC17, CLCab, Hip (Hip1 and Hip1R) or control siRNA. Migration across the wound was imaged in the presence of medium containing 1% serum on glass-bottomed plates using live-cell time-lapse microscopy. (a) Representative HeLa cell trajectories at end time points (24 h) are shown. The MtrackJ plugin of ImageJ was used to manually trace migratory cell tracks, marked in colour. (b) Quantitative analysis of HeLa cell relative net displacement (net displacement from the origin relative to control; left) and average speed (distance migrated per min relative to control; right) were quantified from migratory tracks (mean±s.e.m. of at least 230 cells analysed from 11 independent experiments; as in a. *P<0.05, ***P<0.005, one-way analysis of variance (ANOVA) followed by Newman–Keuls post hoc test). (c) Representative immunoblots of siRNA treatments of HeLa cells as in a. (d) H1299 cell trajectories at end time points (15 h) are shown. The MtrackJ plugin of ImageJ was used to manually trace cell tracks as in a. (e) Representative immunoblots of siRNA treatments as in d. (f) Quantitative analysis of H1299 cell relative net displacement (left) and average speed (right) from migratory tracks (mean±s.e.m. of at least 100 cells analysed from five independent experiments; as in d. *P<0.05, **P<0.01, ***P<0.005, one-way ANOVA followed by Newman–Keuls post hoc test). Migration positions of molecular mass markers are indicated in kDa at the right of the immunoblots shown. Scale bars, 100 μm.