Fig. 1: NHSL3 regulates single and collective cell migration.
From: NHSL3 controls single and collective cell migration through two distinct mechanisms
a MCF10A cells are transfected with pools of control (CTRL) or NHSL3 siRNAs and analysed by Western blots using NHSL3 and GAPDH antibodies. Cells are tracked for 6.5 h and migration persistence is extracted from trajectories of single cells. n = 60 cells, mean ± SEM. b NHSL3 KO clones or parental MCF10A cells are analysed by Western blots using NHSL3 and GAPDH antibodies. Cells are tracked for 6.5 h and migration persistence is extracted from trajectories of single cells. n = 60 cells. c hTERT-HME1 cells are transfected with pools of control (CTRL) or NHSL3 siRNAs and analysed by Western blots. Cells are tracked for 8 h and migration persistence is extracted from trajectories of single cells. n = 60 cells, mean ± SEM. d hTERT-HME1 cells are stained with DAPI (nuclear DNA), phalloidin (filamentous actin) and antibodies targeting cortactin or NHSL3. Single confocal section, scale bar: 20 μm. Overlap of cortactin and NHSL3 over multiple line scans registered to the cell edge. Data are shown as mean ± SEM, n = 15 line scans. e Parental MCF10A (Par) and NHSL3 KO cells are stained with DAPI, phalloidin and NHSL3 antibodies. Single confocal section, scale bar: 10 μm. Overlap of actin and NHSL3 over multiple line scans registered to the cell-cell junction. Data are shown as mean ± SEM, n = 15 line scans. f Migration of monolayers into the wound that is created by lifting an insert is imaged by phase contrast over time and analysed by Particle Image Velocimetry (PIV). Heat maps of the speed of collective parental MCF10A or NHSL3 KO cells display the front edge at the top and time of insert lifting at the left and its evolution over time and space (average of 12 measures, i.e. 3 biological replicates each containing 4 fields of view). Quantification of cell speed across the monolayer at 8 h. Statistical significance is calculated using custom-made R programmes for single cell migration and one-way ANOVA for collective migration. p-values are indicated. Three biological repeats for each experiment gave similar results. Source data are provided as a Source Data file.
