Generation and characterization of CaMKII-TDP-43 Tg mice. (A) Physical map of the CaMKII-TDP-43 fragment for pronuclei injection. The orientation of transcription is indicated by the arrow. The positions of the short hybrid intron derived from an adenovirus splice donor, an immunoglobulin G splice acceptor, and the SV40 poly(A) addition sequence (pA) are indicated. The approximate locations of the Southern blotting and PCR probes are also indicated. Tg mice were identified by the presence of the 4.4-kb KpnI fragment on the Southern blot and the 523-bp PCR band on gel. The restriction sites on the map are as follows: K, KpnI; E, EcoRV; N, NotI; S, SfiI. (B) Genotyping of the Tg mice. The data from PCR (top) and Southern blotting (bottom) analysis of the tail DNAs are exemplified. NT represents the nontransgenic samples. (C) Western blotting of the protein extracts from the hippocampus, cortex, cerebellum, and spinal cord of the WT, NT, and Tg mice, respectively. (D) In situ hybridization patterns of TDP-43 transcripts in the brains of WT and TDP-43 Tg mice (Tg). Bars, 500 µm. (E) Immunostaining patterns of TDP-43 protein in the brains of WT and Tg mice. CA1, CA1 layer; CA3, CA3 layer; DG, dentate gyrus. Bars, 200 µm. Results in B–E are representative of three independent experiments.