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. Author manuscript; available in PMC: 2018 Sep 13.
Published in final edited form as: Neuron. 2017 Sep 13;95(6):1292–1305.e5. doi: 10.1016/j.neuron.2017.08.039

Figure 4.

Figure 4

QAGR RAN protein accumulation in human DM2 autopsy brains, identification of hnRNP A1 as CAGG RNA-binding protein and cellular toxicity assays. (A) IHC showing QAGR immunostaining in frontal cortex, basal ganglia and hippocampus in DM2 but not normal control autopsy tissue. (B) IHC showing QAGR immunostaining (left), hematoxylin and eosin (H&E) staining (middle panels), luxol fast blue (LFB) staining of white matter (right panels). (C) UV crosslinking assay of HEK293T cell extracts with CCUG and CAGG RNA probe. CCUG46, but not CCUG5, forms two predominant RNA-protein complexes. Both CAGG5 and CAGG46 also form two predominant RNA-protein complexes. (D) Coupled Crosslinking and Immunoprecipitation show that CCUG46-protein complexes contain MBNL1 and CAGG46-protein complexes contain hnRNP A1. (E) Cell death measured by lactate dehydrogenase (LDH) assay and cell viability (MTT) assays of T98 cells transfected with vectors expressing CCTG repeats or alternative codons encoding LPAC (red) and vectors carrying CAGG repeats or alternative codon repeats encoding QAGR (blue). Bars show average ± SEM, n = 10). (asterisks: *p<0.0125, **p ≤ 0.0025, ***p ≤ 0.00025). See also Figure S4 and S5.