Fig. 5

RASSF8 mediates the roles of miR-224 in cell growth and invasion. a The cells transfected with RASSF8-overexpressing plasmid or control were subjected to Western blot to determine RASSF8 expression. Colony formation assay was performed in the above cells to detect the roles of RASSF8 in cell proliferation. b Cell invasion assay: The above cells were (2.5 × 104 cells) seeded in serum-free medium in a Matrigel-coated chamber, while the lower chamber was incubated with 10%-FBS containing medium. The cells were invaded for 20 h. Non-invading cells were scraped with a cotton tips and invading cells were fixed and stained with crystal violet, followed by counted. c–d The cells were co-transfected with miR-224 ASO and RASSF8 siRNA under hypoxia, together with controls, and then subjected to colony formation assay (c) and cell invasion assay (d). Western blot assay showed that expression of RASSF8 in the above cells. *P < 0.05, **P < 0.01