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. Author manuscript; available in PMC: 2009 May 1.
Published in final edited form as: Am J Physiol Lung Cell Mol Physiol. 2008 Feb 22;294(5):L902–L911. doi: 10.1152/ajplung.00278.2007

Fig. 11. 20-HETE increases Rac1/2 in BPAECs.

Fig. 11

Lysates of BPAEC 80–95% confluence in passages 4 and 5 were separated using an epitope protein that binds to the PAK-PBD domain (Rac effector protein, p21 activated kinase 1-p21 binding domain) to capture activated Rac associated with GTP. After capture, proteins in Western blots were probed with a primary polyclonal antibody to Rac1/2. The positive control for these experiments is histidine-tagged recombinant Rac1. To determine the effect of 20-HETE on total Rac1/2, crude cell lysates from cells treated identically to those used for capture of activated Rac were probed with the primary antibody to Rac1/2 provided in the kit. A representative image from these experiments appears here. Exposure to 20-HETE increased activated Rac1/2 in a manner that peaked within 10 min (n = 4 isolates of cells studied through 30 min; *P < 0.05 relative to vehicle control), and values remained elevated above vehicle control for 30 min. Total Rac1/2 is not affected by 20-HETE treatment in this time frame.