HGAL modulates IL-6 effects on cell migration: wound assay in HeLa-HGAL cells. Wound assays were performed as described in “Chemotaxis and wound assays.” (A) IL-6 effect on HeLa cell migration in wound assay. (B) IL-6 effect on migration of HeLa-HGAL and HeLA-Neo cells. HGAL expression significantly decreased migration of the cells. (C) siRNA knock-down of HGAL in the HeLa-HGAL cells markedly reversed the HGAL-mediated, IL-6–induced inhibition of cell migration. (D) Western blot analysis of HGAL protein expression in HeLa-HGAL cells following siRNA knockdown. (E) IL-6 effect on migration of HeLa-HGAL mutants. Left panel exhibits results of a representative wound assay in HeLa-HGAL Y128F and control cells. Right panel summarizes the effect of IL-6 on migration of HeLa-HGAL-Y128F, HGAL-128F, and HGAL-ΔC mutants. P value represents difference between HeLa-HGAL Y128F and control cells. Means and standard deviations of 3 independent experiments are demonstrated. See “Immunofluorescence microscopy” for image acquisition information.