Role of S100A10 in Matrigel invasion and cell migration. WT and S100A10−/− (KO) peritoneal macrophages (1 × 105 cells) were added to the upper chamber of Matrigel (MTG)-coated chambers (BD Biocoat chambers, 8-μm pore) and incubated in the presence or absence of Plg (0.5μM). The lower chamber contained MCP-1 or C5a (10 ng/mL). Cells were incubated at 37°C for 48 hours (A). Invading cells were quantified as described in “Matrigel invasion and cell migration.” Data are expressed as mean number of cells per 40× field plus or minus SD of 3 independent experiments. Statistical analysis was performed by Student t test: **P < .01. No significant difference in invasion was observed for WT and S100A10−/− macrophages in the absence of Plg (P < .053) by t test. WT (1 × 105 cells; B) and S100A10−/− macrophages (3 × 105 cells; C) were incubated with Plg, and aprotinin, ϵ-ACA, or CpB, and invasion determined in response to MCP-1 (10 ng/mL) in the lower chamber (B-C). The invading cells were quantified as described in panel A. Statistical analysis was performed by one-way analysis of variance comparison of untreated WT macrophage invasion (B) or untreated S100A10−/− macrophage invasion (C). WT and S100A10−/− macrophages (1 × 105 cells) were added with Plg to the upper chamber of Matrigel-coated chambers with plasmin (0.43 CTA U/mL) or thioglycollate (4%) as chemoattractant in the lower chamber (D). Invasion was quantified as described in panel A. Statistical analysis was performed by Student t test: **P < .01. No significant difference in invasion was observed between WT and S100A10−/− macrophages in the absence of a chemoattractant or when thioglycollate was used as a chemoattractant. WT and S100A10−/− peritoneal macrophages (1 × 105 cells) were added to uncoated invasion chambers in the presence of 0.5μM Plg (E). Cells were incubated at 37°C for 48 hours. Migrating cells were quantified as described in panel A. Statistical analysis was performed by Student t test: **P < .01. No significant difference was observed between WT and S100A10−/− macrophage migration in the absence of Matrigel: ***P < .001. ns indicates not significant.
