Fig. 4

RNF144A-AS1 required miR-30c-2-3p to perform its function. A Venn diagram showing potential binding miRNAs for RNF144A-AS1. B Expression of miR-30c-2-3p in 386 GC tissues and 41 normal gastric tissues from TCGA database. C qRT-PCR analysis shown that the expression of miR-30c-2-3p was negatively regulated by RNF144A-AS1. D Construction of luciferase reporter vectors that contain RNF144A-AS1 sequences with or without mutated miR-30c-2-3p binding sites. E Luciferase activity was measured in HEK293T cells co-transfected with increasing concentration of miR-30c-2-3p mimics and indicated vectors. F qRT-PCR analysis of immunoprecipitated RNF144A-AS1 transcripts in Ago2 relative to IgG immunoprecipitates. G Fold enrichment of miR-30c-2-3p and RNF144A-AS1 in immunoprecipitates from cells treated with miRNA inhibitor or control. H Expression of miR-30c-2-3p in GC cells transfected with miRNA mimics or control. I Tube-formation ability of HUVECs co-cultured with indicated conditioned medium. Scale bars = 200 μm. J, K Effects on cellular migration and invasion by transfecting miR-30c-2-3p mimics or control into MKN45 (J) and AGS (K) cells. Scale bars = 100 μm. L, M Effects of co-transfection of miR-30c-2-3p mimics and RNF144A-AS1 vector on cellular migration (L) and tube-formation (M). Error bars, mean ± SD from triplicate samples. *P < 0.05, **P < 0.01, ***P < 0.001 by Student’s t‐test unless otherwise specified