FIGURE 5.

Generation of transgenic mice with SMC-targeted USP20 expression. A, the USP20 transgenes comprised (from 5′ to 3′) a 481-base pair portion of the SM22α promoter (−440 to +41 relative to transcription start), an N-terminal HA epitope, the 2751-base pair murine USP20 coding sequence, and the bovine growth hormone poly(A) signal. The two catalytic domain mutations contained in the DN-USP20 mutant are depicted as Xs: C154S and H645Q. B, 30 μg of aorta extracts from congenic C57BL/6 mice with SM22α-driven expression of USP20, DN-USP20, or no transgene were immunoblotted (IB) with IgGs specific for the indicated proteins. Shown is an immunoblot from a single experiment representative of three such experiments. C, littermate control and Tg aorta sections were stained with anti-HA (green), Cy3-conjugated anti-SMC actin (red), and DRAQ5 for DNA (blue). Sections were imaged with a ×40 oil objective of a LSM 510 Meta confocal microscope. Scale bars = 50 μm.