Fig. 3
From: An engineered miRNA PS-OMe miR130 inhibits acute lung injury by targeting eCIRP in sepsis
PS-OMe miR130 acts to decrease binding of eCIRP to TLR4. A Biotinylated TLR4 was individually immobilized on a sensor chip SA, and eCIRP (500 nM) incubated with various doses of PS-OMe miR130 (1–50 nM) in PBS was injected as the analyte. SPR analysis demonstrates a dose related decrease in binding affinity of eCIRP for TLR4 when combined with increasing doses of the engineered miRNA. The KD decreased from 6.97 × 10−8 M at 0 nM of the miRNA to 1.39 × 10−7 M at 5 nM (n = 3 independent SPR experiments). B Biotinylated TLR4/MD2 was individually immobilized on a sensor chip SA, and eCIRP (1000 nM) incubated with various doses of the PS-OMe miR130 (1–50 nM) in PBS was injected as the analyte. SPR analysis demonstrates decreased binding affinity of eCIRP for the TLR4/MD2 complex when combined with increasing doses of PS-OMe miR130. The KD decreased from 2.0 × 10−8 M with 0 nM of the miRNA to 1.13 × 10−7 with 5 nM of PS-OMe miR130 (n = 3 independent SPR experiments). C Computational modeling was performed between eCIRP and TLR4 and demonstrated that a strong binding complex is created. D Modeling was performed between a combined eCIRP/PS-OMe miR130 and TLR4 and demonstrates a less stable complex