Figure 7.

Lphn3-induced delay in cell-autonomous and non-cell-autonomous polarized migration is unperturbed by cancer-related GAIN domain mutations in wound healing assays. (a) Representative merged images from bright field and epifluorescence microscopy of mVenus-tagged Lphn3-expressing cells or mVenus control cells depicting wound closure assays captured at the initial scratch into the cell monolayer (zero hours) and at the same scratch area 48 h later, in the absence (IgC-Fc) and presence of receptor ligand Flrt3^ECD-Fc. The yellow dotted lines represent the edges of the scratch at time zero hours. Cells incubated with 10% FBS were used as positive migration controls. Scale bar: 200 µm. (b) Percentage scratch closure data in the presence of IgC-Fc or Flrt3^ECD-Fc represented as the percentage of the total scratch area at zero hours that is occupied by cells at 48h, in bright field images. (c) Ratio between fluorescence area occupied by cells expressing mVenus signal in the wound (mVenus-tagged receptors or control cells) and total fluorescence area in the field of view. The dotted line represents the data corresponding to positive control migration values. Subdomain A (GAIN Sub A) and B (GAIN Sub B) of GAIN domain. mVenus-tagged receptor constructs are indicated by a star (★). Data are represented as the mean values of at least three independent experiments (n = 3). Statistical analysis was performed using one-way ANOVA. Error bars indicate S.E.M., p values between mVenus-tagged Lphn3 variants and control data are indicated by # inside histograms: #### p ≤ 0.0001, ### p ≤ 0.001, ## p ≤ 0.01, # p ≤ 0.05.