Figure 8 | Scientific Reports

Figure 8

From: Dimethyl fumarate blocks pro-inflammatory cytokine production via inhibition of TLR induced M1 and K63 ubiquitin chain formation

Figure 8

Effect of DMF on LPS stimulated polyubiquitin chain formation in RAW264.7 macrophages. RAW264.7 cells were incubated in the presence or absence of 50 μM DMF for 4 h.

Cells were then either left unstimulated or treated with 100 ng/ml LPS for a further 30 min. Cells were lysed and lysates were blotted for the phospho and total proteins indicated (A). Alternatively, Halo-NEMO beads were used to pulldown M1 polyubiquitin chains. Pull downs were then immunoblotted with antibodies specific for M1 chains, K63 chains, IRAK1 or IRAK2. Total ubiquitin was pulled down using Halo-TUBE beads and blotted for the presence of K48 chains (B).

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