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. 2015 Nov 13;10(11):e0142387. doi: 10.1371/journal.pone.0142387

Fig 5. HOXB1 is a direct target of miR-3175.

Fig 5

(A) Diagram of the seed sequence of the miR-3175-binding site within the wild-type HOXB1 3′-UTR and the design of the mutated HOXB1 3′-UTR sequence. The mutated HOXB1 3′-UTR sequence is labeled in red. (B) Luciferase assay of U87 cells after cotransfection with the wild-type or mutant HOXB1 3′-UTR and miR-3175 mimics or miR-3175 mimics NC. The pRL-TK Renilla luciferase reporter vector was used as the internal control. Luciferase activity = firefly luciferase/Renilla luciferase. (C) HOXB1 mRNA expression is modulated by miR-3175. Glioma cell lines were transfected with miR-3175 mimics, miR-3175 inhibitor and the corresponding NCs, and incubated for 72 h. Cells were harvested and assayed using qRT-PCR. (D-E) Western blot analysis of HOXB1 protein levels in glioma cell lines after transfection with miR-3175 mimics, miR-3175 inhibitor and the corresponding NCs at 72 h. (F) The correlation between HOXB1 and miR-3175 expression in human glioma tissue specimens was analyzed with Spearman’s rank correlation. All results are representative of three independent experiments, and a statistical analysis is performed (mean ± SD, *P < 0.05, **P < 0.01, and ***P < 0.001).