Figure 7

Validation of the role of polyamines and ODC1 on the proliferation, invasion, and migration of medulloblastoma. (A) Expression of ODC1 in medulloblastoma cell lines (Daoy and D283 Med) against human normal brain glial cells (HEB) was detected by WB and qPCR. (B) The specific siRNA downregulated the protein and mRNA level of ODC1. (C,D) The effect of ODC1 on the migration and invasion ability of Daoy cells was investigated by transwell assay. Representative images are shown on the left; scale bar: 200 μm. Quantitative analysis of cell migration and invasion levels was shown in the right panel. (E) Effects of ODC1 knockdown and addition of polyamines or DFMO on cell viability were determined by CCK-8 assay. (F) Proliferation ability of Daoy cells after ODC1 knockdown and polyamine supplementation was examined by colony formation assay. The left panel shows the overall colony formation in the entire culture dish. The right panel shows the number of cell colonies in each dish. (G) PBMC were co-cultured with D283 Med cells by transwell assay (0.4 μm) after labeling with cell tracker Red, and the number of PBMC cells chemotactic was observed under fluorescence microscopy.