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. 2000 May 23;97(11):5779–5783. doi: 10.1073/pnas.97.11.5779

Figure 5.

Figure 5

GST pulldown and far Western detection of the 50-kDa 12(S)-HETE binding protein. (Left) ATP-treated cytosol was incubated with GST/ΔSRC-11–212 (lanes 1 and 2) or GST/ΔSRC-11–1,138 (lanes 3 and 4) immobilized on GSH-agarose in the presence or absence of 1 nM 12(S)-HETE. (Right) The 50-kDa 12(S)-HETE binding protein (FPLC fraction 16) was incubated with GST/ΔSRC-11–212 (lanes 5 and 6) or GST/ΔSRC-11–1,138 (lanes 7 and 8) immobilized on GSH-agarose in the presence or absence of 1 μM 12(S)-HETE. Interacting proteins were electrotransferred to nitrocellulose membranes and detected by incubating with 32P-labeled GST/ΔSRC-11–1,138 in the presence of 5 nM 12(S)-HETE for ATP-treated cytosol and 1 μM 12(S)-HETE for 50-kDa 12(S)-HETE binding protein.