Figure 4
Focal adhesion in iPSC derived cranial neural crest. (a–e) Immunofluorescence staining for DAPI (blue) and focal adhesion marker vinculin (green) in iPSC derived CNCC plated into fibronectin coated culture plates. Cells were imaged using confocal microscopy. Scale bar is 50 μm. (f) Quantification and size distribution of FA spots. N > 400 per genotype. An increased labelling of focal adhesion spots in both Q152*KI/WT and Q152*KI/Q152*KI mutant lines is observed compared to their isogenic control WT/WT with no difference between Q152*KI/WT and Q152*KI/Q152*KI (Kruskal–Wallis test, P < 0.0001, P < 0.0001, P > 0.9999, respectively). Less labelling of focal adhesion spots and a decrease in spot size for the N68S/N68S mutant is observed compared to its isogenic control N68SIC/N68SIC (Mann–Whitney test; P < 0.0001). (g) Quantification of number of FA spots per cell. N = > 37 per genotype. An increase in focal adhesion spots in both Q152*KI/WT and Q152*KI/Q152*KI mutant lines is observed compared to their isogenic control WT/WT with no difference between Q152*KI/WT and Q152*KI/Q152*KI (One-way ANOVA, P < 0.0001, P < 0.0001, P > 0.9999, respectively). Fewer focal adhesion spots for the N68S/N68S mutant is observed compared to its isogenic control N68SIC/N68SIC (Unpaired t test; P = 0.0026).
