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. 2015 Mar 24;10(3):e0120583. doi: 10.1371/journal.pone.0120583

Fig 3. Nickel and cobalt activate HEK293 cells via human TLR4 and require MD-2 for activation.

Fig 3

(A) HEK293/hTLR4 cells were transfected with the luciferase reporter plasmids with or without the plasmid encoding hMD-2. After stimulation, the luciferase activity was measured. (B) Mouse TLR4 with either mouse or human MD-2 does not support activation by nickel or cobalt. HEK293/mTLR4 cells were transfected with the luciferase reporter plasmids with or without plasmid encoding MD-2. (C) Mutation of histidines at positions 456 and 458 in hTLR4 nearly abolishes responsiveness to nickel and cobalt ions. HEK293 cells were transfected with luciferase reporter plasmids, plasmid encoding hMD-2 and plasmid encoding either wild type or mutant hTLR4. Luciferase activity was measured as indicated in Methods. #p≥0,01 (not significant); *p<0,01; **p<0,001; ***p<0,0001 (t-test, compared to the unstimulated control). The chart legend applies to all three panels.