Fig. S10.

Interaction of Gα₁₃ with RIP3 in platelets stimulated with thrombin. Washed platelets (3 × 10⁸ /mL) from WT mice were stimulated with 0.006 U/mL thrombin for the indicated times at 37 °C under stirring. The platelets were lysed with equal volumes of 2× Nonidet P-40 lysis buffer containing protease inhibitor mixture tablets on ice for 30 min. After centrifugation at 17,000 × g and 4 °C for 10 min, the supernatants were immunoprecipitated with anti-Gα₁₃ antibody or IgG (control) overnight. After incubation with Protein A/G Plus Agarose beads at 4 °C for 2 h, the proteins were analyzed by Western blot analysis with anti-RIP3 and anti-Gα₁₃ antibodies. The figures are representative of three independent experiments.