Figure 3. Ribitol and ribose increase the levels of ribitol-5-P and cytidine diphosphate (CDP)-ribitol in patient-specific Walker-Warburg syndrome (WWS) myotubes.
(A) Detection of ribitol, ribose, ribitol-5-P, and CDP-ribitol in WWS FP4 myotubes that had been treated or not with ribitol or ribose for 5 days (n = 3 for each cohort). Error bars represent standard errors of three independent experiments. (B–C) Sorbinil treatment for 5 days results in decreased α-dystroglycan (α-DG) functional glycosylation, as shown by western blot for IIH6 in wild type (WT) myotubes (B). β-DG was used as loading control. (C) Bar graph shows quantification of IIH6 (from B) normalized to β-DG and shown as the fold difference of WT. Error bars represent standard errors of three independent experiments. (D–E) Sorbinil treatment counteracts the positive effect of ribose on α-DG functional glycosylation. (D) Representative western blot shows reduction of IIH6 staining in FP4 myotubes that had been treated with both ribose and sorbinil. β-DG was used as loading control. (E) Bar graph shows quantification of IIH6 (from D) normalized to β-DG and shown as the fold difference of FP4 + ribose. Error bars represent standard errors of five independent experiments. Significance was evaluated by the one-way ANOVA followed by the Sidak’s multiple comparison test in (A) and the unpaired Student’s t test in (C and E). *p<0.05, **p<0.01, ***p<0.001.
Figure 3—figure supplement 1. Liquid chromatography with tandem mass spectrometry (LC/MS-MS) standard curves.
