Fig. 5

a Sequence alignment of DANCR with potential wild type and mutant type of miR-335-5p (wt-DANCR-335 and mut-DANCR-335) and miR-1972 (wt-DANCR-1972 and mut-DANCR-1972) targeting sites. b-c Luciferase reporter assay was applied to verify the targeted binding effect between DANCR 3′UTR and miR-335-5p or miR-1972, **, P < 0.01, as compared and normalize to mimic control group, respectively. d-e RIP binding assay was performed using input from cell lysate, normal mouse IgG or anti-Ago2. Relative expression levels of DANCR and miR-335-5p or miR-1972 were determined by qRT-PCR. **P < 0.01 vs anti-normal IgG group. f-i A re-executed CCK-8 and EDU assay presented that promotion of miR-335-5p or miR-1972 (transfection of miR-335-5p mimics or miR-1972 mimics) led to significantly suppression of proliferation, while the suppressive effect was reversed by wt-DANCR but not be done by mut-DANCR-335 or mut-DANCR-1972. j-k Re-executed transwell assay also demonstrated that it was wt-DANCR that can attenuate miR-335-5p- or miR-1972-induced inhibition of metastasis, when the theoretical binding sites DANCR might provide for miR-335-5p or miR-1972 were mutated, the reversing effect vanished. For F-K, data were normalized to NC mimic group, and **P < 0.01, & P > 0.05 as comparing with miR-335-5p mimics group or miR-1972 mimics group, respectively. Data were shown as mean ± SD from three independent experiments