Figure 3: Hip-binding by CLC is required to rescue the migration defect induced by CLC depletion. | Nature Communications

Figure 3: Hip-binding by CLC is required to rescue the migration defect induced by CLC depletion.

From: Clathrin light chains are required for the gyrating-clathrin recycling pathway and thereby promote cell migration

Figure 3

Wound-healing assays were performed in H1299 clones that stably expressed either empty vector, siRNA-resistant WT CLCa, siRNA-resistant WT CLCb, siRNA-resistant mutant CLCa or siRNA-resistant mutant CLCb after 72 h of treatment with siRNA targeting CLCab to deplete endogenous CLCs. Mutant (mut) CLCs are defective for Hip-binding. Migration across the wound was imaged in the presence of medium containing 1% serum using live-cell time-lapse microscopy on plastic tissue culture plates for 10 h. (a) Representative cell trajectories at end time points are shown. The MtrackJ plugin of ImageJ was used to manually trace cell tracks, marked in colour. Scale bars, 250 μm. (b) Quantitative analysis of relative net displacement (left) and relative average speed (right; mean±s.e.m. of at least 40 cells analysed from two independent experiments; as in a. **P<0.01, two-way analysis of variance followed by Bonferroni post hoc test). (c) Representative immunoblots of siRNA treatments as in a. Migration positions of molecular mass markers are indicated in kDa at the right of the immunoblots shown.

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