Fig. 4 | Communications Biology

Fig. 4

From: Wnt5a is a TLR2/4-ligand that induces tolerance in human myeloid cells

Fig. 4

Wnt-proteins interact with TLR proteins. a (upper panel) Wnt5a and Wnt3a bind to TLR4, TLR4/MD2, and TLR2. SPR sensorgrams obtained after injection of Wnt protein at 100 nM over TLR4, TLR4/MD2 or TLR2 immobilized in Fc 2, 3, and 4, respectively, are shown. The responses are expressed as resonance units (RU) after subtraction of the sample buffer response in the reference surface (Fc 1). The increase in response at the time of sample injection is a bulk effect caused by traces of CHAPS in the sample solution. (lower panel) Steady-state response (Req), calculated after kinetic evaluation of sensorgrams using a 1:1 model, plotted vs. analyte concentration. The binding curves obtained after injection of Wnt5a and Wnt3a at 25, 50, and 100 nM over TLR4, TLR4/MD2 and TLR2 are shown from left to right in the lower panel. Curves were fit to a one-site hyperbola model for calculation of steady-state affinity and maximal binding. The traces are a representative of two experiments. The results are summarized in Table 1. b Wnt5a interacts directly with TLR4. In vitro binding assay for detection of the rWnt5a and rhTLR4-his interaction. The protein band that is only present when both proteins are mixed, and that is specifically recognized by anti-Wnt5a and anti-His antibodies (arrow), contains Wnt5a and TLR4 proteins (as determined by LC-MS/MS; LC-MS/MS data are shown in Supplementary Table 3). Stars, protein bands detected by anti-Wnt5a detects in the well only containing rWnt5a

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