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. 2016 Jul 28;8(60):100975–100988. doi: 10.18632/oncotarget.10900

Figure 3. SATB2 is a direct target of miR-449a.

Figure 3

(A) The predicted binding sites of miR-449a in the SATB2 3′UTR. (B) Luciferase activity of the SATB2 3′UTR reporter in 293T cells transfected with 100 nmol/L miR-NC or miR-449a mimics, 200 ng of the psiCHECK-SATB2 3′UTR reporter plasmid, SATB2-3′UTR-1 (binding site 1), SATB2-3′UTR-1-MUT (binding site 1 mutant), SATB2-3′UTR-2 (binding site 2), SATB2-3′UTR-2-MUT (binding site 2 mutant) or the control reporter plasmid for 24 hours. (C) miR-449a, SATB2 mRNA expression levels in different CRC cell lines using the CCD-112CoN cell line as the control. (D) SATB2 protein levels in Lovo, HT29, HCT116 and 293T cells treated with 100 nmol/L miR-NC or miR-449a mimics. (E) Effect of time-dependent inhibition of miR-449a mimics (upper panels) on the SATB2 protein (lower panels) in HCT116 cells. (F) Effect of dose-dependent inhibition of miR-449a mimics (upper panels) on the SATB2 protein (lower panels) in HCT116 cells. (G) Immunohistochemistry results (upper panels) and western blotting results for SATB2 in xenograft tumors derived from HCT116 cells stably expressing miR-449a or the negative control.