Expression of A20 in the ascending aorta and aortic arch of diabetic ApoE-null mice prevents the development of atherosclerotic lesions by inhibiting PKCβII phosphorylation and blunting the induction of RAGE.

<p>(A) Transgene expression was confirmed by X-gal staining in rAd.βgal-transduced vessels 5 days following transgene delivery (n = 3 mice/group) and demonstrate the expression of the transgene in medial SMC (M) as well as the adventitia at the level of the aortic root, albeit not in all cells. Image amplification 100× and 400×. (B) A20 expression was verified by real time RT-PCR in two rAd.A20-transduced vessels, using human A20 specific primers that do not recognize mouse A20. Our data indicate significant expression of human A20 in aortic roots of rAd.A20 but not saline treated mice. Results are shown as average± SE. (C) H&E stained aortic root sections at the level of the first coronary from 20 week-old diabetic ApoE-null mice treated with saline, rAd.A20 or rAd.βgal. Images are shown at 100× and 400× as indicated by the scale bar. The asterisk indicates the level of the first coronary branch, Arrows define the intima (I) and the media (M). ApoE-competent, non-diabetic C57BL/6 and non-diabetic ApoE-null mice were used as controls. Blood glucose and cholesterol levels (cholest) are listed below the sections. *P<0.05 compared to saline, ** P<0.01 compared to rAd. βGal. Data shown are representative of 4 to 6 mice per group. I/M ratios were calculated after analysis of 10 serial sections per vessel. (D) phospho-(p)PKCβII (5 days) and RAGE (14 days) immunostaining in aortic arches 5 and 14 days after transgene delivery. Data shown in C are representative of all sections analyzed (n = 3 mice per group, 2–3 sections analyzed per vessel). Image amplification 200×.