Lower A20 protein levels contrast with increased mRNA levels in failed vein grafts of diabetic patients.

Abstract

<p>(A) A20 protein expression and (B) mRNA levels (real-time PCR, lower panel) in failed vein grafts recovered from non-diabetic (ND) and patients with type I and type II DM (t1 and t2 DM) show lower A20 protein levels in failed vein grafts of diabetic patients as compared to non-diabetic patients, despite significantly higher A20 mRNA levels in these samples. Expression of GAPDH was checked as a loading control and to quantify relative A20 expression by densitometry. A20/GAPDH ratios are listed below the WB. Data shown are representative of 4 patients in each group. Samples were all run on the same gel but are shown as separate because they were not loaded contiguously. (B) A20 mRNA levels of the same failed vein grafts shown in 4A demonstrate increased A20 mRNA levels in failed vein grafts from diabetic vs. non-diabetic patients. 18S ribosomal RNA was used to normalize the data. Data is expressed as fold increase of A20 mRNA using the A20 mRNA level detected in the failed vein graft of the non-diabetic patient as a baseline value. (C) A20 mRNA levels analyzed by real time PCR from failed vein grafts of diabetic (DM) and non-diabetic (ND) patients show significantly higher A20 mRNA in failed vein grafts of diabetic versus non-diabetic patients; **p = 0.0016. 18S ribosomal RNA was used to normalize the data. Natural log transformed data (ln) are presented as mean±SEM of 6 failed vein grafts per group. The 6 failed vein grafts from diabetic patients included 2 type I and 4 type II DM patients. Data is expressed as fold increase of A20 mRNA using the A20 mRNA level detected in the failed vein graft of the non-diabetic patient shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0014240#pone-0014240-g001" target="_blank">Figure 1A</a>, as a baseline value.</p