Fig 1: VWMD cell lines with EIF2B2 mutations have normal translation activity in unstressed conditions. (A) Equal numbers of control and VWMD patient cell lines were fed 35S-labeled met and cys, lysed and proteins separated on 4%–12% NuPAGE SDS PAGE gels and detected by phosphorimaging. The intensity of each lane was quantified and the averages ±SD of the lane intensity of each VWMD patient sample relative to the matched control from three independent experiments is shown below a representative gel. (B) The levels of EIF2B1, EIF2B2, EIF2B3, EIF2B4, and EIF2B5 proteins in VWMD patient and control lymphoblasts was assessed by western blotting. Brackets enclose blots for which the indicated EIF2B protein was stripped and reprobed for GAPDH as a loading control. Quantification shows the average relative EIF2B protein levels from three independent experiments ±SEM below a representative blot. (C) Phospho-eIF2a levels were determined by western blotting in VWMD patient and matched controls. The average abundance of phospho-eIF2a (“EIF2a-pS51”) relative to total EIF2a from three independent experiments ±SEM is shown below a representative blot. Student's t-test was used to assess significance in VWMD samples versus control for all experiments.