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

Figure 10. Recombinant STC1 inhibits thrombin-mediated ERK1/2 activation through suppression of superoxide production.

Figure 10

A549 cells were pre-treated with the following agents, either singly, or in combination, followed by thrombin treatment (1 U/mL) for 10 min at 37 °C: paraquat (40 μM) for 4 h; MnTBAP (0.1 mM) for 30 min; STC1 (200 ng/mL) for 2 h. 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). (C) A549 cells, treated as above were incubated with MitoSOX (5 μM) and Hoechst (0.2 μg/ml) for 10 min, and MitoSOX fluorescence was visualized (excitation 510 nm, emission 580 nm). 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).