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. 2004 Jan 8;23(2):322–332. doi: 10.1038/sj.emboj.7600044

Figure 5.

Figure 5

Effect of lipid raft disruption and zinc depletion on TNFα-induced JNK and IκB phosphorylation. (A) Zinc depletion inhibits TNFα-induced JNK activation but not IκB phosphorylation and degradation. HeLa cells were cultured in 1% FBS/DMEM for 1 h and further incubated in the presence of MCD or TPEN for 30 min before exposure to TNFα treatment in the same medium. Protein samples were extracted in TNE buffer (containing phosphatase inhibitor) and subjected to immunoblot analysis with anti-phospho-JNK, anti-phospho-IκB and control antibodies. (B) Rapid disruption of membrane rafts activates JNK and IKK signaling pathways. NIH 3T3 cells were cultured in 1% FBS/DMEM for 1 h and further incubated in the presence of MCD or TPEN for 30 min before exposure to TNFα treatment in the same medium. Protein samples were extracted in TNE buffer (containing phosphatase inhibitor) and subjected to immunoblot analysis with anti-phospho-JNK, anti-phospho-IκB and control antibodies as indicated.