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Fig. 2 | Cancer Cell International

Fig. 2

From: The lncRNA NEAT1 promotes the epithelial-mesenchymal transition and metastasis of osteosarcoma cells by sponging miR-483 to upregulate STAT3 expression

Fig. 2

miR-483 targets STAT3 and reduces its expression. a The potential binding site for miR-483 in the 3′UTR of STAT3 according to the bioinformatics prediction from the Starbase database. b U2OS and MG-63 cells were transfected with a luciferase plasmid containing the wild type 3′UTR or mutant 3′UTR of STAT3 along with miR-483 mimics or mimics NC. The interaction between miR-483 and the 3′UTR of STAT3 was examined using a dual luciferase assay. ce U2OS and MG-63 cells were un-transfected or transfected with inhibitor NC, miR-483 inhibitor, mimics NC, or miR-483 mimics. c The relative expression of miR-483 in control, inhibitor NC- or miR-483 inhibitor-transfected U2OS and MG-63 cells was measured using qRT-PCR. The expression of miR-483 was normalized to U6. d The relative expression of STAT3 in U2OS and MG-63 cells was also measured using qRT-PCR. e The levels and phosphorylation of STAT3 in U2OS and MG-63 cells were examined using western blotting. GAPDH served as the input control. All experiments reported in this study were performed at least three times, which referred to biological replicates. Each biological replicate consisted of 3 wells with cells of the same passage, which referred to technical replicates. Error bars denoted mean ± SD. p values were calculated using unpaired two-tailed student’s t-test (b) or one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test (ce). **p < 0.01 and *p < 0.05

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