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. Author manuscript; available in PMC: 2009 Mar 9.
Published in final edited form as: J Biol Chem. 2006 Sep 18;281(45):34705–34715. doi: 10.1074/jbc.M605738200

FIGURE 3. TNF-α induces recruitment of signaling adaptors and RhoA into lipid rafts.

FIGURE 3

Human airway smooth muscle cells were extracted with cold Triton X-100 and fractionated by sucrose density gradient centrifugation (A) or stimulated with TNF-α for the indicated times (B) before isolation of Triton X-100-soluble (S) and Triton X-100-insoluble, octyl glucoside-soluble raft fractions (R). Equal volume samples were separated by SDS-PAGE and analyzed with specific antibodies for the expression of TNFR1, signaling adaptors, signaling intermediates, and RhoA as indicated. Lipid rafts were identified by the expression of the raft-resident protein, caveolin-1, whereas soluble fractions expressed RhoGDI. The cytoskeletal protein, smooth muscle actin (sm-actin), was restricted to soluble fractions.