Fig. 4

Overexpression of circ_001422 promotes the proliferation and metastasis of OS cells. a & b. EdU incorporation (scale bar, 100 μm) and colony formation assays were performed to evaluate the effect of circ_001422 overexpression on the cell proliferation ability. c & d. Flow cytometry was performed to evaluate the effect of circ_001422 overexpression on the cell cycle distribution and apoptosis. e. Transwell migration and Matrigel invasion assays were performed to evaluate the effect of circ_001422 overexpression on cell migration and invasion. Scale bar, 100 μm. f. Western blot analysis was performed to evaluate the effect of circ_001422 overexpression on the expression of cell cycle-, apoptosis-, and EMT-associated marker proteins. g. Representative images of xenograft tumors in nude mice after subcutaneous implantation of 143B cells stably transduced with circ_001422 overexpression or control lentiviral vectors. h & i. Effect of circ_001422 overexpression on tumor growth and tumor weight in nude mice. j & k. Representative images of lung tissues and lung sections stained with H&E in the lung metastasis mouse model. Scale bar, 1000 μm. l. Tumor tissues were sectioned and subjected to immunohistochemical staining for Ki-67, PCNA, E-cadherin, N-cadherin and Vimentin. Scale bar, 100 μm. n.s., not significant; ^*P < 0.05; ^**P < 0.01; ^***P < 0.001