Generation of a Trim3 knockout mouse and validation of TRIM3 depletion.

(A) Schematic representation of the Trim3 gene and the Trim3 targeting construct. Homologous recombination of the targeting construct produces a mutant Trim3 gene containing a Neo^r cassette flanked by frt sites (grey boxes, f) and loxP sites (white boxes, l) flanking exons 3-5. Recombination of the loxP sites results in excision of exons 3-5; recombination of the frt sites results in excision of the Neo^r cassette. Forward primers (a-d), reverse primers (1-4) and BglII restriction sites (▲) used for genotyping are indicated (see Figure S1C). (B) Western blotting confirms the absence of TRIM3 in hippocampus (Hc), cerebellum (Cb) and cortex (Cx). (C) Immunocytochemistry confirms the absence of TRIM3 in cultured hippocampal neurons derived from Trim3 knockout mice (DIV12). Control: Synaptic vesicle protein SV2. (Scale bars: 20 µm.).