FIG. 7.

SPRC induced the nuclear translocation of STAT3, followed by transcriptional activation. (A) HUV-EC-C were treated with SPRC (100 μM) or interferon (IFN)-γ (100 ng/ml) for 45 min, and then, cytoplasmic and nuclear proteins were isolated for Western blotting. GAPDH and histone H3 were used as a loading control for cytoplasmic and nuclear proteins, respectively. (B) Primary HUVEC were treated with SPRC (100 μM) for 45 min and then stained with specific antibody (red) or DAPI (blue). Scale bar, 20 μm. (C) HUV-EC-C were treated with SPRC (100 μM), IFN-γ (100 ng/ml), or WP1066 (10 μM) for 45 min. The nuclear extract was isolated and used for electrophoretic mobility shift assay. The shift band of STAT3 is indicated with a solid arrowhead. IFN-γ was used as a positive control, and WP1066 was used to inhibit STAT3. (D) HUV-EC-C were treated with SPRC (100 μM) for 45 min. The nuclear extract was isolated and used to perform chromatin immunoprecipitation with an STAT3 antibody. The change of downstream promoters was analyzed by real-time polymerase chain reaction. ^#p<0.01 between groups, as indicated. All experiments were repeated at least thrice. To see this illustration in color, the reader is referred to the web version of this article at www.liebertpub.com/ars