Figure 4
Figure 4. DNMT activities and protein levels. HUVECs were treated with 50 μM DL-Hcy or 1 mM AZC in control medium for 48 hours. Nuclear extracts (20 μg) were prepared and incubated with hemimethylated double-stranded DNA for DNMT1 activity (A) or with unmethylated double-stranded DNA for DNMT3 activity (B) in the presence of S-adenosyl-L-[methyl-3H] methionine. DNMT activities were determined by the radioactivity level of DNA substrates. Values represent mean (± SD) from 3 separate experiments with triplicated samples (n = 9). *P < .01 versus control. Protein levels of DNMT1 and DNMT3 were examined by Western blotting with antibodies against DNMT1 or DNMT3, respectively, and reblotted with β-actin antibody.

DNMT activities and protein levels. HUVECs were treated with 50 μM DL-Hcy or 1 mM AZC in control medium for 48 hours. Nuclear extracts (20 μg) were prepared and incubated with hemimethylated double-stranded DNA for DNMT1 activity (A) or with unmethylated double-stranded DNA for DNMT3 activity (B) in the presence of S-adenosyl-L-[methyl-3H] methionine. DNMT activities were determined by the radioactivity level of DNA substrates. Values represent mean (± SD) from 3 separate experiments with triplicated samples (n = 9). *P < .01 versus control. Protein levels of DNMT1 and DNMT3 were examined by Western blotting with antibodies against DNMT1 or DNMT3, respectively, and reblotted with β-actin antibody.

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