<p>Aspirin–treated, washed platelets were stimulated with 2MeSADP (100 nM) at various time points (A) or with varying concentrations of 2MeSADP (B) for one minute under stirring conditions at 37°C. The lysates were then subjected to western blotting analysis and probed with anti-phospho-(Y416) and total c-Src antibodies as lane loading control. Washed murine (WT or c-Src KO) platelets, without aspirin-treatment, were stimulated with 2MeSADP (100 nM) for 3.5 minutes and TXB<sub>2</sub> levels were analyzed (C) as described for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0016586#pone-0016586-g001" target="_blank">Figure 1</a>. The data are represented as the % Fold increase over the control. Aspirin-treated washed platelets were stimulated with 2MeSADP (100 nM) for (30–120 seconds) or convulxin (100 ng/ml) for 30 seconds under stirring conditions at 37°C (D). The lysates were then subjected to western blotting analysis and probed with anti- phospho- Syk(Y 525/526) and total Syk antibodies as lane loading control. The data are representative of at least 3 separate experiments.</p
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