Fig. 4
miR-224 mimics facilitates the proliferation, invasion and migration of GC cells by targeting the 3′-UTR of Smad4 and p21 directly. a qRT-PCR analysis of miR-224 levels in BGC-823 and SGC-7901 cells transfected with miR-224 mimics or the control mimics. b EdU analysis of the cell proliferation ability in BGC-823 and SGC-7901 cells transfected with miR-224 mimics or the control mimics. Scale bar: 20 μm. c Statistical analysis of the EdU-positive cell ratio in transfected GC cells. d and e CCK-8 analysis of the cell proliferation ability in BGC-823 (d) and SGC-7901 (e) cells transfected with miR-224 mimics or the control mimics. f The scratch wound healing assays in BGC-823 and SGC-7901 cells transfected with miR-224 mimics or the control mimics. Scale bar: 500 μm. g Statistical analysis of the cell migration in the scratch wound healing assays. h Transwell invasion and migration assay in BGC-823 and SGC-7901 cells transfected with miR-224 mimics or the control mimics. Scale bar: 100 μm. i Statistical analysis of the cell numbers passing through the transwell chamber in the transfected BGC-823 and SGC-7901 cells. All datas were the means ± SD. j Western blot analysis of the protein expression level of PTEN, HOXD10, p21 and Smad4 in transfected GC cells. k qRT-PCR analysis of the mRNA expression level of p21 in transfected GC cells. l qRT-PCR analysis of the mRNA expression level of Smad4 in transfected GC cells. m The pMIR-Smad4-WT or pMIR-Smad4-Mut reporter construct was cotransfected with control or miR-224 mimics into BGC-823 and SGC-7901 cells, and the dual luciferase activity was determined at 48 h after transfection. n The pMIR-p21-WT or pMIR-p21-Mut reporter construct was cotransfected with control or miR-224 mimics into BGC-823 and SGC-7901 cells, and the dual luciferase activity was determined at 48 h after transfection. All datas were the means ± SD. o Western blot analysis of Smad4 and p21 protein level in BGC-823 cells transfected with hsa_circ_0004872 siRNAs. p Western blot analysis of Smad4 and p21 protein level in GC cells transfected with hsa_circ_0004872 overexpression vector (circ_4872) or the mutated hsa_circ_0004872 overexpression vector (circ_4872-Mut)