Fig 1: Calgranulin activation of toll-like receptor 4 (TLR4) is shared across amniotes. NF-κB activity for species-matched TLR4/MD-2/CD14 complexes in the presence of calgranulins from (A) human, (B) mouse, (C) opossum, and (D) chicken. A “+” in the table below each series indicates which components are included in that treatment. Lipopolysaccharide (LPS) is used as a positive control for expression and activation of the complex. Polymyxin B is included to control for endotoxin-mediated activation of the complex. Activity is normalized to LPS activity of positive control within each biological replicate (i.e., for human, each technical replicate is divided by the average LPS activation of hTLR4, hMD2, and hCD14 for that biological replicate). Points are the technical triplicates from three biological replicates. Bold lines are the mean of the biological replicates. Error bars are SEM of the biological replicates. A two-tailed t-test was used to assess significance of difference in mean between the indicated series (**p-value <0.01).
Fig 2: Lipopolysaccharide (LPS) and calgranulin exhibit different complex requirements. (A–C) show NF-κB activation of mouse toll-like receptor 4 (TLR4) with different combinations of mouse and opossum MD-2 and CD14. Activation by: (A) LPS, (B) mouse S100A9, and (C) opossum S100A9. (D–F) show NF-κB activation of human TLR4 with different combinations of human and mouse MD-2 and CD14. Activation by: (D) LPS, (E) human S100A9, and (F) mouse S100A9. Letters indicate which species the component was taken from: human (H), mouse (M), or opossum (O). NF-κB is normalized to LPS activation of the control complex for that species. Points are the technical triplicates from three biological replicates. Bold lines are the mean of the biological replicates. Error bars are SEM of the biological replicates. A two-tailed t-test was used to assess significance of difference in mean between the indicated series (*p-value <0.05, **p-value <0.01).
Supplier Page from DNASU for LY96 (Homo sapiens) in pLX304 (Gateway V5-tagged lentiviral expression vector)