Figure 6.

Induction of STAT3 in iNOS-null mice at 3 hr after PH occurs normally, when compared with controls. (A) STAT3 DNA binding activity was determined by electrophoretic mobility-shift assay (EMSA) and supershift assays at various time points after PH. Maximal STAT3 binding activity occurred at 3 hr post-PH. This representative EMSA depicts STAT3 binding activity at baseline (time 0) and 3 hr after PH in controls and iNOS-null animals. Lanes 1 a-c contain time 0 extracts from control animals with added preimmune sera (a), total STAT3 antibody (b), or phosphorylated STAT3 antibody (c). Lanes 2 a-c contain extracts from controls at 3 hr post-PH in a similar order; lanes 3 a-c and 4 a-c contain extracts from iNOS-null animals at time 0 and 3 hr post-PH, respectively. (B) Total and phosphorylated STAT3 protein levels were determined by Western blot assay in control and iNOS-null mice at baseline (time 0), 30 min, and 3 hr after PH. Lanes 1 and 2 represent two individual animals used for the experiment at each time point.