Figure 6
Figure 6. Brg1 is a direct target of miR-139-5p. (A) Schematic representation of luciferase constructs used for reporter assay. The alignment among the binding regions of the miR-139-5p within the Brg1 3′UTR of human and mouse, mutated derivative of Brg1 3′UTR and the paring sites of miR-139-5p, and miR-139-5p sequence is shown. (B) Reporter analysis on HeLa cells transfected with constructs described in the figure. Data represent mean ± SD (N = 3). (C) western blotting of Brg1 expression in miR-139-5p–transfected HeLa (endogenous miR-139-5p expression is not detected) and PD31 (endogenous expression of miR-139-5p is detected) cells. (D) Locked nucleic acid-modified miR-139-5p inhibitor was transfected in PD31 cells, and lysates were prepared for immunoblotting with the indicated antibodies. Data are representative of 2 independent experiments. (E) Intracellular staining of Brg1 expression in LSK cells from reconstituted mice expressing miR-139-5p-RV (left). Bar graph represents relative mean fluorescence intensity (MFI) of Brg1 (N = 3, right). (F) Intracellular staining of Brg1 expression in LSK cells from miR-139-5p–knockdown mice. Bar graph represents relative MFI of Brg1 (N = 3, right).

Brg1 is a direct target of miR-139-5p. (A) Schematic representation of luciferase constructs used for reporter assay. The alignment among the binding regions of the miR-139-5p within the Brg1 3′UTR of human and mouse, mutated derivative of Brg1 3′UTR and the paring sites of miR-139-5p, and miR-139-5p sequence is shown. (B) Reporter analysis on HeLa cells transfected with constructs described in the figure. Data represent mean ± SD (N = 3). (C) western blotting of Brg1 expression in miR-139-5p–transfected HeLa (endogenous miR-139-5p expression is not detected) and PD31 (endogenous expression of miR-139-5p is detected) cells. (D) Locked nucleic acid-modified miR-139-5p inhibitor was transfected in PD31 cells, and lysates were prepared for immunoblotting with the indicated antibodies. Data are representative of 2 independent experiments. (E) Intracellular staining of Brg1 expression in LSK cells from reconstituted mice expressing miR-139-5p-RV (left). Bar graph represents relative mean fluorescence intensity (MFI) of Brg1 (N = 3, right). (F) Intracellular staining of Brg1 expression in LSK cells from miR-139-5p–knockdown mice. Bar graph represents relative MFI of Brg1 (N = 3, right).

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