Fig 1: Blocking NMD increases Grn mRNA in GrnR493X mouse model and GRN+/R493X patient fibroblasts. (A) Wild-type and GrnR493X/R493X MEFs were treated for 5 h with 50 μM cycloheximide, 10 μM MG-132, or 10 μg/mL tunicamycin. Gene expression was determined by qPCR. (B) Wild-type and GrnR493X/R493X mice (n = 4–7 per group) were acutely injected with 20 mg/kg body weight cycloheximide dissolved in PBS. After 4 h, gene expression was determined by qPCR. (C and D) Upf1 and Upf2 were knocked down in wild-type, Grn+/R493X, and GrnR493X/R493X MEFs using DharmaFECT1. After 3 d, gene expression was determined by qPCR (C), and protein levels were determined by Western blot analyses (D). (E) UPF1 and UPF2 were knocked down in control and GRN+/R493X HDFs using DharmaFECT1. After 3 d, gene expression was determined by qPCR. All data are presented as mean ± SD; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, as determined by one-way ANOVA followed by Tukey’s post hoc test (A), or two-way ANOVA followed by Bonferroni’s post hoc test (B, C, and E). Asterisk in D indicates a nonspecific band. n.s., not significant.