Fig. 5: The SUI1 domain of eIF-2D is required for poly-GA production. | Nature Communications

Fig. 5: The SUI1 domain of eIF-2D is required for poly-GA production.

From: A C. elegans model of C9orf72-associated ALS/FTD uncovers a conserved role for eIF2D in RAN translation

Fig. 5

a Schematic of the eif-2D genetic locus. Missense mutations (gk904876, gk561128) and CRISPR/Cas9-engineered alleles (syb3364[eif-2D::GFP::3xFLAG], syb3432[eif-2D ΔSUI1]) are shown. The PUA and SUI1 domain are indicated with brackets. b An endogenous reporter allele (syb3364) shows cytoplasmic eif-2D expression. Dark oval or circular shapes of various sizes are nuclei of different cell types. Representative images of the head of two animals (L4 stage) are shown. Negative control is a wild-type (N2) animal (L4). Scale bar = 10 μm. c, f Worm lysates were processed for western blotting and immunostained with poly-GA and α-tubulin antibodies. d, g Quantification of poly-GA expression on western blots. The experiments were repeated four times. Mean ± s.e.m. One-way ANOVA with Dunnett’s multiple comparisons test was performed. e, h The G4C2 repeat RNA with C9orf72 intronic RNA, and act-1 mRNAs were assessed by RT-PCR. The experiments were repeated four times in e and five times in h. Mean ± s.e.m. One-way ANOVA with Dunnett’s multiple comparisons test was performed.

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