Fig. 3
From: Defects in t6A tRNA modification due to GON7 and YRDC mutations lead to Galloway-Mowat syndrome
Effects of YRDC and GON7 mutations on t6A biosynthesis. a Evaluation of fitness of Δsua5 yeast strains expressing human YRDC variants (spots are from 10-fold serial dilutions of cell suspensions at OD600nm = 0.5, and three independent clones were evaluated) and b western blot analysis on total protein extracts from Δsua5 yeast cells expressing human YRDC variants using anti-hYRDC antibody. c, d Mass spectrometry (LC-MS/MS) quantification of t6A modification in total tRNAs extracted from Δsua5 yeast cells expressing human YRDC variants (c) (mean ± s.e.m. of two independent LC-MS/MS experiments (technical replicates), each measuring samples from three independent yeast transformants; one-way ANOVA (F (5,28) = 269.4, p < 0.0001), Dunnett’s multiple comparisons test, ***p = 0.0008, ****p < 0.0001) and from cultured primary skin fibroblasts from controls (two unaffected individuals) and affected individuals with either the p.Tyr7* GON7 mutation (two individuals), or the YRDC mutations (three individuals) or with the p.Arg352Gln OSGEP mutation in the homozygous state (individual «CP» described in Braun et al.13) (d) (mean ± s.e.m. of two independent LC-MS/MS experiments (technical replicates), each measuring samples from three independent cell culture experiments; one-way ANOVA (F (3,44) = 6.446, p < 0.001), Dunnett’s multiple comparisons test, n.s. = 0.4894, *p = 0.0169, ***p = 0.008). Source data are provided as a Source Data file
