Generation of a <i>Trim3</i> knockout mouse and validation of TRIM3 depletion.

<p>(A) Schematic representation of the <i>Trim3</i> gene and the <i>Trim3</i> targeting construct. Homologous recombination of the targeting construct produces a mutant <i>Trim3</i> 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 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075603#pone.0075603.s001" target="_blank">Figure S1C</a>). (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 <i>Trim3</i> knockout mice (DIV12). Control: Synaptic vesicle protein SV2. (Scale bars: 20 µm.).