Fig. 3

Galectin-9 medium (Gal-9 M) induces angiogenesis and acute inflammation in vivo. a Matrigel plug angiogenesis assay with PBS negative control and Gal-9 M (139.5 nmol/L). Matrigel plugs containing Gal-9 M had significantly more hemoglobin than the PBS control group (p < 0.01). n Number of mice per group. b and c Cryosection of Matrigel plugs stained for von Willebrand factor (vWF) displayed a significant increase in blood vessel formation in response to Gal-9 M compared with PBS, a negative control (p < 0.001). d Matrigel plug assay with Janus kinase (Jnk) and p38 signaling inhibitors. Jnk inhibitor significantly reduced Gal-9 M-stimulated angiogenesis in the Matrigel plugs, whereas p38 inhibitor did not. n Number of mice per group (p < 0.02). e Gal-9 M induces in vitro monocyte (MN) chemotaxis. MN chemotaxis assays were performed using a modified Boyden chamber. MN migration was determined in response to Gal-9 M. PBS and N-formylmethionyl-leucyl-phenylalanine (fMLP) were used as negative and positive controls, respectively. Three high-power fields (HPFs) were counted per well, and the assay was performed in quadruplicate. n Number of replicates. Means are presented with SEM, and p values < 0.05 were considered significant. f Acute inflammatory arthritis in mouse knees injected with Gal-9 M. Results were measured as knee circumference from the day of injection (day 0). Gal-9 M at concentrations of 139.5 nmol/L and 1.39 μmol/L in 20 μl of PBS induced significantly higher inflammation. At 139.5 nmol/L, knee circumference was increased by ~ 3-fold (p < 0.0068), whereas at 1.39 μmol/L, the increase in knee circumference was 2-fold (p < 0.01). g F4/80 staining of mouse knee cryosections injected with Gal-9 M and PBS. Knees were snap-frozen and cryosectioned, then stained with 4′,6-diamidino-2-phenylindole (DAPI; blue) and F4/80 (red). Gal-9 M-injected knees had increased MN/macrophage staining compared with those injected with PBS