Generation and validation of <i>Vcpkmt</i> knockout.

<p>(<b>A</b>) Schematic overview of K315 trimethylation of VCP monomer by VCPKMT. AdoMet = S-adenosyl methionine (<b>B</b>) Targeting of the <i>Vcpkmt</i> gene. Diagram shows the endogenous murine <i>Vcpkmt</i> locus, the position of the targeted gene and the recombined gene. (<b>C</b>) Representative genotyping result from <i>Vcpkmt</i> wild-type (+/+), knockout (-/-) and heterozygous (+/-) mice. (<b>D</b>) Immunostaining for VCPKMT in wild-type and <i>Vcpkmt</i>^<i>-/-</i> mouse embryonic fibroblasts. DNA counterstain with DAPI. Scale bars 10 μm. (<b>E</b>) The gene expression of <i>Vcpkmt</i> relative to <i>Gapdh</i> in various murine tissues measured with quantitative RT-PCR. n = 3 mice, means ± SD.