Fig. 2
miR-190 induced BC cell growth by down-regulating p27 and promoting G1/S phase transition. a–d The indicated cells were seeded into 6-well plates and cultured to 70–80% confluence; after synchronization in 0.1% FBS medium for 24 h., cells were cultured in complete medium for another 24 h. and then subjected to cell cycle analysis by flow cytometry as described in Materials and Methods. e GO analysis of potential miR-190 targeted genes. f, g, h, k Cell lysates from the indicated cells were evaluated for p27, cyclin D1, CKD4, and CKD6 expression via western blots. β-Actin or GAPDH served as the loading control. (i, j, l, m) A soft agar assay was used to determine the effect of p27 overexpression (i, j) or knockdown (l, m) on anchorage-independent growth compared with vector control cells; Representative images of colonies from the indicated cells were captured under microscopy after 3 weeks of incubation. The number of colonies was counted under microscopy and the number of colonies was counted and presented as colonies per 104 cells. The symbol (*) indicates a significant difference (*P < 0.05, **P < 0.01, ***P < 0.001; #P < 0.05, ##P < 0.01, ###P < 0.001)