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Fig. 3 Pathogenic variants in PRMT9 affect the cilia length of skin fibroblasts from affected individuals (A) Fibroblasts from control and individuals’ cells in ciliary conditions were stained with an antibody against acetylated α-tubulin (cilia, red). Nuclei were stained with DAPI (blue). (B) Based on 20 fields in three independent experiments (100–150 cells per experiment), mean percentages of ciliated cells of unaltered individual cells compared to control cells are shown in a histogram. The control is the mean of three independent controls, and error bars represent standard deviation. (C) Individual cells present longer cilia. Data are presented as a scatterplot with a line indicating the mean value and error bars indicating the standard deviation. The control is the mean of three independent controls. Statistical significance was determined using the unpaired non-parametric Mann-Whitney test (n = 300; ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). (D) Assessment of Hh-signal transduction by quantification of the expression levels of two Hh target genes (PTCH1 and GLI1) on Hh-stimulated cells (+SAG) from controls and the individual with (−FCS) and without (+FCS) inducing ciliogenesis. All the results are representative of three different experiments. Data are presented as relative expression levels ± SEM. p values were calculated using a two-way ANOVA test (Tukey’s multiple comparison test). ns, not significant; ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.