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. 2020 Feb 21;21(4):e47852. doi: 10.15252/embr.201947852

Figure 1. Physical interaction of Gal‐3 and Gal‐1 with CC and CXC chemokines.

Figure 1

  • A–E
    Chemokine–galectin interactions were detected by using a solid‐phase immunoassay. For this, 46 human CC and CXC chemokines were adsorbed on nitrocellulose membranes and the stripes incubated in parallel with (A) TBS or (B) TBS containing biotinylated galectins (the representative image shows a processed membrane tested with labeled Gal‐3). Signals had been generated by using SA‐HRP and chemiluminescence reagents. (C, D) The blots were subjected to densitometric analysis, and (E) all independent experiments were combined (binding chemokines in light blue, Gal‐3: n = 5, Gal‐3 CRD: n = 5, Gal‐1: n = 4).
  • F–H
    For SPR‐based experiments, (F) Gal‐3 (density 650 RU), (G) Gal‐3 CRD (density 1180 RU), and (H) Gal‐1 (density 130 RU) were immobilized and increasing concentrations of CXCL12 were passed over the flow cells. The red curve represents a single‐site fit to the data. Insets are representative sensorgrams of CXCL12 testing on immobilized galectin. Data represent the mean ± SD from six (F) or three (G and H) independent experiments.