Figure 3.

Neointimal hyperplasia triggers phosphorylation of UPS20 on Ser334. Carotid arteries of female mice from Figure 2 were serially sectioned and immunostained with IgGs specific for USP20 phosphorylated on Ser334 (“p-USP20”), total USP20, ACTA2, or no protein (isotype control, “Ctrl”), and counterstained for nuclear DNA (Hoechst 33342). Confocal fluorescence photomicrographs are shown from uninjured (native) carotids (A) and contralateral carotids subjected to endothelial denudation (“injured,” B); scale bars = 20 μm. Dotted lines denote the external elastic lamina. C and D, for each carotid artery section, protein immunofluorescence in the tunica media and intima was normalized to DNA fluorescence. These ratios were normalized to the mean value obtained for WT carotid arteries and plotted for individual mice (along with means ± SD) from three native carotids (C) and six injured carotids (D) from each genetic group. Compared with WT: ∗p < 0.01. L, lumen; USP20, ubiquitin-specific peptidase 20.