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. 2009 Jul 1;297(3):F662–F670. doi: 10.1152/ajprenal.00146.2009

Fig. 3.

Fig. 3.

Activation of Raf/MEK/ERK pathway by 20-HETE and stable 20-HETE mimetics. Representative Western blot demonstrates activation of Raf/MEK/ERK pathway using specific antibodies. Cell lysates were obtained from LLC-PK1 cells incubated in serum-free media containing 20 μM 20-HETE, 20 μM 5,14-20-HEDGE (HEDGE), and 20 μM 5,14-20-HEDE for 3 h. Indomethacin (2 μM) was added to prevent COX-mediated metabolism 60 min before 20-HETE and agonist incubation. Changes in levels of p-ERK1/2, phosphorylated MEK1/2 (p-MEK1/2), and phosphorylated Raf-1 (p-Raf-1) in the presence of 20-HETE, 5,14-20-HEDGE, and 5,14-20-HEDE are shown. β-Actin was used as a loading control. Immunoblots represent 2–8 results for each antibody group.