Figure 1.

β5i Expression Was Upregulated in Ang II-Infused Mice and in Patients with HR

(A) WT mice were infused with Ang II at a dose of 3,000 ng/kg/min or with saline for 3 weeks. We then measured proteasome (caspase-like, trypsin-like, and chymotrypsin-like) activities in the mouse retinas (n = 3 per group). (B) qPCR analysis of expression of proteasome catalytic subunits (β1, β2, β5, β1i, β2i, and β5i) in the retinas (n = 4 per group). (C) Immunoblotting analysis of immunosubunits (β1i, β2i, and β5i; upper) and quantification of protein levels (lower; n = 3 per group). We used glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as an internal control. (D) Immunostaining for β5i with anti-β5i antibody in retinal sections (left) and quantification of red fluorescence intensity (right; n = 5 per group). (E) Immunostaining for β5i antibody in retinal tissues from normal human controls and from HR patients (left), and quantification of red fluorescence intensity (right; n = 3 per group). Nuclei were counterstained with DAPI (blue). Scale bars: 50 μm. (F and G) Analysis by ELISA of β5i concentration (F) and chymotrypsin-like activity (G) in serum of controls (n = 31) and of HR patients (n = 30). Data are presented as mean ± SEM. *p < 0.05, **p < 0.01 versus saline or normal controls.