Fig. 3
Effect of TSP2 protein expression on the migration and proliferation of glioma cells. a–b Western blot bands (upper panels) showing TSP1 and TSP4 protein expression in the C6WT, C6TSP2+/+, and C6TSP2−/− groups. Experiments were repeated 3 times with cells at different passages. Densitometry analysis of Western blot bands (lower panels) showed that the TSP1 (C6WT vs. C6TSP2+/+: P = 0.248, C6WT vs. C6TSP2−/−: P = 0.344, C6TSP2+/+ vs. C6TSP2−/−: P = 0.067) and TSP4 (C6WT vs. C6TSP2+/+: P = 0.999, C6WT vs. C6TSP2−/−: P = 0.469, C6TSP2+/+ vs. C6TSP2−/−: P = 0.448) protein levels were not significantly different between the C6WT, C6TSP2+/+, and C6TSP2−/− groups. c In the CCK-8 assay, the optical density values at 450 nm were recorded to determine the proliferation rates of C6WT, C6TSP2+/+, and C6TSP2−/− cells after 24, 48, or 72 h of cultivation. Compared with C6WT cells, the proliferation of C6TSP2+/+ cells was increased (P < 0.001), but the proliferation of C6TSP2−/− cells was decreased (P < 0.001). d Representative images of scratch wounds of C6WT, C6TSP2+/+, and C6TSP2−/− cells after 24 h (bar: 500 μm). e Ratio of the unhealed area between 0 and 24 h after wounding in C6WT, C6TSP2+/+, C6TSP2−/− cells. C6TSP2+/+ cells (P < 0.001) migrated faster, and C6TSP2−/− cells (P < 0.001) migrated slower than C6WT cells at 24 h. f–g In the CCK-8 assay, the optical density values at 450 nm showed that the proliferation rates of C6TSP2−/− cells and C6TSP2+/+ cells treated with the exogenous TSP2 protein were not significantly different from those were not treated with the exogenous TSP2 protein after 24, 48, and 72 h of culture (P = 0.174 and P = 0.551, respectively). h Representative images of scratch wounds of C6 cells after 48 h (bar: 500 μm). i Ratio of the unhealed area between 0 and 48 h after wounding in C6TSP2−/− cells treated with or without the exogenous TSP2 protein. The group treated without the exogenous TSP2 protein was regarded as the control group. C6TSP2−/− cells (P = 0.002) treated with the exogenous TSP2 protein migrated faster at 48 h than those that were not treated with the exogenous TSP2 protein. j Representative images showing the migration of C6TSP2−/− cells cultured with or without the exogenous TSP2 protein in the chamber after 24 h (bar: 100 μm). k The number of migrated C6TSP2−/− cells treated with the TSP2 protein was greater than the numbers of cells in the other groups after 24 h (P < 0.001)