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. 2016 Aug 12;8(33):54136–54148. doi: 10.18632/oncotarget.11250

Figure 6. Knockdown of fractalkine inhibits cell migration ability and lung metastasis in osteosarcoma.

Figure 6

A. Total protein was collected from MG63 cells stably expressing fractalkine shRNA. A vector-only control is also shown (control). Western blotting was used to assess the expression of fractalkine and ICAM-1. Actin was used as the loading control. B. The cell migration ability of MG63 cells stably expressing a shRNA vector and control vector was measured using the Transwell assay. C. The protein levels of p-PI3K, PI3K, p-Akt and Akt in control-shRNA and fractalkine-shRNA cells was examined by western blot. D. To induce pulmonary metastases, cells were injected into the mouse tail vein and those mice were sacrificed after 28 days later with developed lung metastatic nodules. Compared to the control mice, there are fewer and smaller tumors which were seen on the lungs of mice injected with osteosarcoma cells transfected with shRNA against fractalkine. E. H&E staining of lung metastatic nodules of MG63 injected mice. The scale bars shown on 100× images correspond to 250 μm. F. The lungs of MG63 injected mice were removed and inflated with 10 % paraformaldehyde fixative. The number of lung metastatic nodules was counted under a dissecting microscope. Results are shown as mean ± SEM. # represents P < 0.01 fort-test comparisons to cells harboring only the empty vector as control.