ATII induces Ets-1 and PDGF-D via H2O2. (A) SMCs were treated with 10– 7M ATII for the times indicated prior to extraction of total RNA and assessment of PDGF-D mRNA expression by RT-PCR. (B) SMCs were treated with 10–7M of ATII for 4 hours and then total cell lysates were assessed for PDGF-D protein expression by Western blotting. Peptide blockade experiments involved incubation of the PDGF-D antibody solution with or without the PDGF-D peptide (sc-23573P; 3.3 μg/mL final) for 1 hour at 37°C prior to incubation with filters. (C) PDGF-D mRNA expression by RT-PCR (left) or real-time PCR (right) in SMCs incubated with ATII for 4 hours and pretreatment with either PEG-catalase (50 U/mL) or the equivalent amount of bovine serum albumin (BSA) for 18 hours. (D) Ets-1 expression in SMCs incubated with ATII for 4 hours and pretreatment with either PEG-catalase (50 U/mL) or the equivalent amount of BSA for 18 hours; ns denotes nonspecific band. The data are representative of at least 2 independent experiments. Error bars represent SEM performed in duplicate or triplicate. *P < .05 relative to control using Student t test.
