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. 2018 May 1;50(5):57. doi: 10.1038/s12276-018-0082-5

Fig. 4. DANCR and HSP27 are both targets of miR-577.

Fig. 4

a DANCR and HSP27 shared the same theoretical miR-577 binding sites, as predicted by targetscan and DIANA-LncBase. b miR-577 was expressed at a lower level in CRC tissues and cell lines than in non-CRC tissues and NCM460, as measured by qRT-PCR. U6 was used as the normalization control. ##P < 0.01 vs. the non-CRC group, **P < 0.01 vs. the NCM 460 group. c miR-577 was up- and down-regulated by transfection of pre-miR-577 or anti-miR-577, as detected by qRT-PCR; &P > 0.05, ***P < 0.001 vs. control group. d HSP27 was negatively regulated by miR-577, as determined by qRT-PCR and western blotting; &P > 0.05, **P < 0.01 vs. the control group (Nos. 1–5 represent the control group, pre-miR-577-NC group, pre-miR-577 group, anti-miR-577-NC group and anti-miR-577 group, respectively). e Diagram of the constructed HSP27 reporter plasmid containing the wild-type or mutant miR-577 binding site. f Co-transfection of pre-miR-577 and pmirGLO-HSP27-wt led to significant weakening of fluorescence compared with the pre-miR-577 and pmirGLO-HSP27-mut co-transfection group, as determined by luciferase assays. &P > 0.05, **P < 0.01 vs. the pre-miR-577-NC group. g Diagram of the constructed DANCR reporter plasmid containing wild-type or mutant miR-577 binding sites. h DANCR was also negatively regulated by miR-577, as determined by qRT-PCR; &P > 0.05, **P < 0.01 vs. the control group. i, j Weakened fluorescence was present in the pre-miR-577 and pmirGLO-HSP27-wt co-transfection groups, as determined by luciferase assays. &P > 0.05, **P < 0.01 vs. the pre-miR-577-NC group. k, l The RIP assay was performed using input from the cell lysate, normal mouse IgG or anti-Ago2. Relative expression levels of DANCR and miR-577 were determined by qRT-PCR. **P < 0.01 vs. the anti-normal IgG group. The data are shown as the means ± SD from three independent experiments