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. 2021 Jul 1;11:708739. doi: 10.3389/fcimb.2021.708739

Figure 4.

Figure 4

Binding of rLIC13086 with extracellular matrix components. (A) Each ECM component was coated onto ELISA plates for 16 h at 4°C and incubated with rLIC13086 (1 μg) for 2 h at 37°C. The detection was performed by incubation with HRP-conjugated anti-HisTag mAb (1:10,000), or anti-rLIC13086 antiserum (1:5,000) and secondary HRP-conjugated anti-mouse IgG (1:5,000). (B) Glycosaminoglycans were immobilized onto ELISA plates for 16 h at 4°C, incubated with rLIC13086 (1 μg) for 2 h at 37°C, following detection by HRP-conjugated anti-HisTag mAb (1:10,000). (C) Coated laminin (1 μg) was incubated with different concentrations of rLIC13086 (0–0.5 μM) for 2 h at 37°C and detected with HRP-conjugated anti-HisTag mAb (1:10,000). (D) Laminin (1 μg) was immobilized onto ELISA plates, incubated with different concentrations of sodium metaperiodate (0–100 mM) and interacted with rLIC13086 (1 μg) at 37°C for 2 h. Detection was performed with HRP-conjugated anti-HisTag mAb (1:10,000). BSA and Fetuin were used as negative controls. The bars represent the means of three different experiments in triplicates with the corresponding standard deviation. * Indicates significant difference calculated by Student t-test with p-values < 0.05.