Fig. 3: Generation of ATXN2 BAC transgenic mice. | Nature Communications

Fig. 3: Generation of ATXN2 BAC transgenic mice.

From: ATAXIN-2 intermediate-length polyglutamine expansions elicit ALS-associated metabolic and immune phenotypes

Fig. 3

A BAC clone containing the human ATXN2 locus with flanking genomic sequence. B, C Western blot for total ATXN2 (mouse and human) in cortical lysates of 6-months-old mice. Black arrowhead, human transgene. White arrowhead, mouse ATXN2. C Quantification of western blots as in B. Each datapoint corresponds to one cortical lysate obtained from one animal (n = 8 for NTg, n = 7 for Q22/+ and n = 6 for Q33/+). D–F Immunohistochemistry for ATXN2 (mouse and human) in 6-months-old mice. G PCR on mouse genomic DNA to confirm ATXN2 CAG repeat integrity. H qRT-PCR to determine ATXN2 transgene copy number. Q22/+ and Q33/+ mice contain six and four copies, respectively. Each datapoint corresponds to one animal (n = 3 for NTg, n = 3 for Q22/+ and n = 4 for Q33/+). I–K Targeted Locus Amplification: high throughput sequencing reads covering the complete transgene sequence (primer set 2). J, K Identification of transgene integration sites. Bar graphs show individual mice and mean ± SD, n ≥ 3 animals per genotype. Scale bar is 40 D, 30 E, and 100 µm F. Source data are provided as a Source Data file for C, H.

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