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. 2011 Nov 18;287(3):2168–2178. doi: 10.1074/jbc.M111.269811

FIGURE 1.

FIGURE 1.

Platelets mediate melanoma cell adhesion in vitro but have no effect on melanoma cell transmigration or proliferation. A, murine endothelial (b.End.3) cells were grown to confluence in transwell chambers. Murine B16 melanoma cells were added to the upper well and allowed to migrate toward the lower well containing buffer, platelets, or SDF-1 (200 ng/ml). B16 melanoma cells overexpressing CXCR-4 or control vector (B16-pLNCX2) were applied. No significant difference was observed regarding B16 transmigration in the absence or presence of platelets. *, p < 0.05 in the presence of SDF-1. B, static adhesion of B16 cells in the presence (PLT) or absence (ctrl.) of immobilized murine platelets (1 × 108/well). Data are representative of four experiments, each with three analyzed wells/condition (means ± S.D.). *, p < 0.05. C, proliferation of B16 cells was measured by [3H]thymidine incorporation after 48 h of co-culture with freshly isolated murine platelets (PLT) (2 × 107/well) at the indicated ratios or with Tyrode's buffer (ctrl.). Data from three experiments, each with three analyzed wells/condition (means ± S.D.), are shown. No significant difference was observed between groups. D, B16-luc tumor cell formation following subcutaneous inoculation of 4 × 105 B16-luc cells in 20 μl of PBS into the left footpad of C57BL/6 mice. Arrows indicate time points of intraperitoneal injection of 200 μl of anti-murine platelet-hyperimmune serum (1:10, closed circles) to deplete (depl.) platelets or control serum (open squares). Tumor volumes (mm3) were measured using a caliper at the indicated time points. Data are from two experiments (means ± S.D.) with 4 to 5 mice per group. No significant difference was observed between groups. E, B16-luc tumor metastasis to draining popliteal lymph nodes (LN). On day 21 following subcutaneous inoculation of B16-luc (4 × 105 cells/20 μl of PBS) into the left footpad of C57BL/6 mice, draining popliteal lymph nodes were processed to determine the metastatic tumor burden by bioluminescence measurements (in relative light units, RLU). Data are means ± S.D. from two experiments with 4 to 5 mice per group. No significant (n.s.) difference was observed between animals after platelet depletion (PLT-depl.) and control mice (ctrl.).