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. 2019 Oct 28;15:343–358. doi: 10.1016/j.omtm.2019.10.008

Figure 5.

Figure 5

Silencing of Mutant Ataxin-3 in SCA3 Knockin Mice

(A) Expression of mature miATXN3 guide strands in the cerebellum after DCN administration. Total RNA was isolated from the cerebellum for small RNA TaqMan. MicroRNA input levels were normalized to U6 small nuclear RNA and set relative to AAV-GFP-treated mice. (B) Lowering of total ATXN3 mRNA in cerebellum of DCN-injected mice. Total RNA was isolated from cerebellum and qRT-PCR was performed to detect the mouse wild-type ATXN3 mRNA. RNA input levels were normalized to GAPDH and set relative to AAV-GFP treated mice. (C) Expression of mature miATXN3 guide strands in the brain stem after cisterna magna administration. Performed as described in (A). (D) Lowering of total ATXN3 mRNA in cerebellum of cisterna magna injected mice. Performed as described in (B). (E) Expression of mature miATXN3 guide strands in the brain stem after cisterna magna administration. Performed as described in (A). (F) Lowering of total ATXN3 mRNA in brain stem of cisterna magna-injected mice. Performed as described in (B). (G) Reduction of mutant ataxin-3 protein in the brain stem after cisterna magna delivery. TR-FRET immunoassay was performed on tissue homogenates to specifically detect the mutant ataxin-3 protein containing more than 37 glutamine repeats (no detection of wild-type mouse ataxin-3). (H) Reduction of mutant ataxin-3 protein in the cerebellum after cisterna magna delivery.