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. 2015 Oct 14;20(1):58–70. doi: 10.1111/jcmm.12702

Figure 5.

Figure 5

Immediate and efficient dissociation of RelA from ubiquitinated IκBα after TNF stimulation requires functional p97/VCP to subsequently allow (i) rapid and efficient proteasomal degradation of ubiquitinated IκBα and (ii) rapid and efficient RelA nuclear translocation. (A) Inhibition of p97/VCP promotes the co‐precipitation of ubiquitinated IκBα with RelA in response to TNF. (B) Inhibition of p97/VCP promotes its association with ubiquitinated proteins but not ubiquitinated IκBα in response to TNF. (C) Inhibition of p97/VCP prevents/decelerates the TNF‐induced liberation of RelA from ubiquitinated IκBα. (A–C) IPs of RelA (A), p97/VCP (B) or IκBα (C) were performed from cytosolic fractions of cells pretreated with NMS‐873 (2.5 μM) for 10 min. prior to TNF (10 ng/ml) stimulation for the indicated times. LMB (10 ng/ml) was applied 15 min. after TNF stimulation. Samples were analysed by IB using the indicated antibodies.