Fig. 2

DARS-AS1 acts as a sponge of miR-628-5p in CC cells. a Subcellular fractionation assay showed the main cytoplasmic location of DARS-AS1 in CC cells. b FISH assay was determined that DARS-AS1 was primarily located in the cytoplasm of CC cells. Scale bar = 10 μm. c Luciferase reporter assay found the specifically decreased luciferase activity of DARS-AS1 under the upregulation of miR-628-5p rather than other miRNAs. d Ago2-RIP assay followed by RT-qPCR confirmed the high enrichment of DARS-AS1 and miR-628-5p in Ago2-assembled RISC. e The successful overexpression of miR-628-5p was tested via RT-qPCR. f The binding sites between wild-type DARS-AS1 and miR-628-5p (DARS-AS1-WT and miR-628-5p-WT) were predicated by ENCORI, and the sequences of mutant DARS-AS1 and miR-628-5p (DARS-AS1-Mut and miR-628-5p-Mut) were designed. g Luciferase reporter assay validated the specific combination between DARS-AS1 and miR-628-5p at the predicted sites. h RT-qPCR verified the evident interference of miR-628-5p in two CC cells by miR-628-5p inhibitor. i CCK-8 assay found that the declined viabilities of cells under DARS-AS1 suppression were recovered after co-transfected with sh-DARS-AS1#1 and miR-628-5p inhibitor. j Colony formation assay assessed that cell proliferation capacities hindered by sh-DARS-AS1#1 was then reversed under the co-inhibition of miR-628-5p. k, l TUNEL assay (scale bar = 100 μm) and flow cytometry analysis unveiled that DARS-AS1 silence-promoted cell apoptosis was cut down in response to further inhibition of miR-628-5p. The data from three independent experiments were presented as mean ± SD. **P < 0.01