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Fig. 5. uPA induces activation of Tyk2 in MCs. (A) Quiescent MCs were treated with 20 nM uPA for the indicated times and phosphorylated Tyk2 protein was visualized in cell lysates by immunoblotting with specific anti-phospho-Tyk2 antibodies (upper panel). MCs incubated in medium without uPA served as a control. The middle panel demonstrates the amount of total Tyk2 loaded on the gel for each sample. Results are representative of three independent experiments. Quantification (mean ± s.e.m.) of the results for these experiments by densitometry is shown below. Significance between control unstimulated and stimulated cells was determined by Student's t-test (*P<0.05). (B) MCs were left uninfected or were infected with wild-type Ad5Tyk2 or with the mutant form, Ad5Tyk2C, and then stimulated for 20 minutes with 20 nM uPA. Phosphorylated Stat3 protein was visualized in cell lysates by immunoblotting with specific anti-(P)-Tyr-Stat3 antibodies. Noninfected MCs incubated in medium without uPA served as a control. The middle panel demonstrates the equal amount of total Stat3 loaded on the gel for each sample. Results are representative of three independent experiments. Quantification (mean ± s.e.m.) of the results by densitometry is shown for three experiments is shown below. Significance between control unstimulated and stimulated cells was determined by Student's t-test (*P<0.05).

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