Effect of lower concentration of homocysteine on the ATF3 gene expression in reporter assay and phosphorylation of JNK/SAPK.

(A) HUVECs transfected with 2 μg of pLuc-1850 reporter plasmid were stimulated with the indicated concentration of homocysteine as in Figure 2B, and ATF3 promoter activity was assayed as described in “Materials and methods.” The results are the average of 3 independent experiments with an SE bar. Basic represents the reporter plasmid without ATF3 gene promoter. Statistically significant fold induction was found in the presence of homocysteine; *P < .05, †P < .01, n = 3. (B) HUVECs were treated with the indicated concentration of homocysteine for 2 hours, and the whole-cell extracts (40 μg) were analyzed by Western blot with anti-JNK/SAPK (JNK) or anti-phospho JNK/SAPK antibody (JNK-○P).