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. 2015 Dec 7;5:18117. doi: 10.1038/srep18117

Figure 9. Recombinant STC1 (rSTC1) inhibits thrombin-induced and PAR1-mediated ERK1/2 activation and superoxide production.

Figure 9

A549 cells were pre-treated with SCH79797 (4 μM) for 30 min or STC1 (200 ng/ml) for 2 h, followed by thrombin treatment (1 U/ml) for 10 min at 37 °C. Cells were harvested, lysed and proteins were resolved on SDS-PAGE. Western blots were reacted with anti-ERK1/2 or anti-pERK1/2. (A) Representative blots are shown. (B) Quantitation of pERK1/2 band intensities normalized to total ERK1/2 bands; bar graph depicts the mean ± SEM (n = 3). Thrombin induces superoxide through PAR1; recombinant STC1 attenuates thrombin-induced superoxide generation. A549 cells were treated as above, incubated with MitoSOX (5 μM) and Hoechst (0.2 μg/ml), and MitoSOX fluorescence was visualized (excitation 510 nm, emission 580 nm). (C) Representative images are shown; magnification: 200X. (D) Quantitation of MitoSOX fluorescence in A549 cells treated as above; bar graph depicts the mean± SEM (n = 3).