Journal: eBioMedicine

Article Title: SLK is mutated in individuals with a neurodevelopmental disorder

doi: 10.1016/j.ebiom.2025.105725

Figure Lengend Snippet: Quantification of wound assay, Golgi appearance and microtubule distribution in patient-derived fibroblasts compared to control . (a) Representative images of sub-confluent fibroblast cells derived from either the index individual in Family 1 (IV:1) or control cell lines at 0 h of wound-induced cell migration assay. Scale bar: 1000 μm. (b) Wound closure was followed serially for 29 h during which a significant difference in cell migration was observed between the two cell lines. Scale bar: 1000 μm. (c) Percent wound closure was evaluated up to 29 h. (d) The remaining area of the wound was measured in μm 2 . The percent wound closure and area of the wound were measured using the ImageJ plugin Wound healing size tool image tool analysis ( https://github.com/AlejandraArnedo/Wound-healing-size-tool/wiki ). Data from 50 cells were collected and analysed. (e) Control cells showing perinuclear compact and polarized Golgi apparatus that are directed toward the migrating wound edge. (f) IV:1-derived fibroblast cells depicting a dispersed and non-polarized (oriented) Golgi apparatus towards the migrating/wound edge. (g) A representative image of normally polarized and parallel orientated microtubules in control cells. (h) A representative image of individual IV:1-derived fibroblast cells showing an un-polarized chaotic microtubule distribution; microtubules labelled with α-tubulin (green) and the centrosome is labelled with Pericentrin (orange) antibodies. (i) The total area of Golgi from 100 cells was measured using the ImageJ software. Error bars represent stdev. (j) The directional orientation of Golgi from 100 cells was assessed and presented as percent mean polarization with error bars representing stdev of 3 replicates. (k) The direction of orientation (paralellness) of microtubules was measured using the LPX ImageJ plugin ( https://lpixel.net/services/research/lpixel-imagej-plugins/ ) ; and data presented as mean and stdev of images of 100 cells. Data from 50 cells were collected and analysed and p-value was calculated based on Welch's t-test for (c, d, i, j and k). The red squares represent datapoints collected from control sample, the green circles represent datapoints collected from the patient sample (IV:1). The horizontal line represents the mean.

Article Snippet: Error bars represent stdev. (j) The directional orientation of Golgi from 100 cells was assessed and presented as percent mean polarization with error bars representing stdev of 3 replicates. (k) The direction of orientation (paralellness) of microtubules was measured using the LPX ImageJ plugin ( https://lpixel.net/services/research/lpixel-imagej-plugins/ ) ; and data presented as mean and stdev of images of 100 cells.

Techniques: Derivative Assay, Control, Cell Migration Assay, Migration, Software