Fig. 5: NHSL3 i2 regulates the speed of follower cells in collective migration through its interaction with MENA/VASP family proteins.
From: NHSL3 controls single and collective cell migration through two distinct mechanisms
a Predicted binding sites on NHSL3 for ABI1, IRSp53 and MENA/VASP are deleted in isoform 2, as indicated (Δ1 to Δ4). b AlphaFold2-generated models of interactions between NHSL3 and partners. Confidence of the interaction is indicated as the AF2 score. c GFP immunoprecipitates from stable rescued KO lines are analysed by Western blots. Two biological repeats with similar results. d Collective migration of NHSL3 KO cells stably expressing the different NHSL3 i2 mutant forms, assessed as in Fig. 1b (average of 12 measures, i.e. 3 biological replicates each containing 4 fields of view). Statistical significance is calculated with Kruskal-Wallis test and p-values are indicated. e MCF10A NHSL3 KO cells stably expressing NHSL3 i2 or NHSL3 i2_Δ4 are allowed to migrate collectively into the wound for 8 h and monolayers are stained with DAPI, phalloidin and VASP antibodies. Scale bar: 10 μm. Intensity of GFP fluorescence, VASP immunofluorescence and phalloidin is plotted as percentages from -2.5 to 2.5 µm across the cell-cell junction (mean ± SEM of 45 measures, i.e. 3 biological replicates each containing 15 line scans). The three biological repeats of all displayed experiments gave similar results. Source data are provided as a Source Data file.
