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Fig. 8
Functional evaluation of DHODH inhibition. A Phenocopy of cartilage-loss phenotype following treatment with 10 µM teriflunomide between 50% epiboly to 5 days post-fertilization, and inhibition of sox10 expression at 24 hours post-fertilization following teriflunomide treatment, shown by in situ hybridisation. B Dose-dependent inhibition of cell numbers in SK-N-SH (n = 6), LAN5 (n = 3), and SK-N-AS (n = 5) cells after 3 days treatment with leflunomide. Data are from CCK8 assays with untreated controls given an arbitrary value of 1. All 100 μM samples are significantly different from untreated samples (p < 0.05). Furthermore, at 100 μM both SK-N-SH and LAN5 are significantly more sensitive than SK-N-AS (LAN5:SK-N-AS, p = 0.025; SK-N-SH:SK-N-AS, p = 0.002; ANOVA with post-hoc Scheffé test). Images show morphology of cells treated with 100 µM leflunomide for 3 days (note no increase in neurite formation, a differentiation marker). C Gene ontology terms statistically significantly enriched within the list of differentially expressed genes (log2-fold change >2, p < 0.05) according to DAVID software analysis. Number of terms within each category is listed. ****p < 5 × 10-5; ***p < 10-4; **p < 10-3; * p < 10-2; no asterisk, p < 0.05.