Cultured smooth muscle cells were prepared from abdominal aortas from 2-month-old WT and Rsk2KO mice, cleaned of adventitia, cut into 1 mm2 pieces, and left undisturbed for 10 days in culture medium. Cells grown in serum-free medium were stimulated with serum in the presence and absence of LJH685 (10 μM), the PDK1 inhibitor GSK2334470 (10 μM), and the ERK1/2 inhibitor U0126 (10 μM) added 4 hours before serum stimulation for 5 min.

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