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. 2017 Apr 1;8(28):45213–45223. doi: 10.18632/oncotarget.16779

Figure 4. MiR-27a antioligonucleotides increase the expression of Apaf-1 in colorectal CSCs.

Figure 4

(A) Apaf-1 mRNA 3′ UTR contained conservative sequence at the miR-27a binding site predicted by the TargetScan database. (B) Expression of Apaf-1 in FHC, HT29 and SW480 CSCs and non-CSCs. (C) Colorectal patients’ tumor and adjacent normal tissues were digested by using collagenase type III to obtain the single cell suspension. After sorting, expression of Apaf-1 in normal colorectal tissue cells, colorectal CSCs and non-CSCs was evaluated by western blot analysis. (D) Luciferase reporter assay in HT29 and SW480 CSCs. The wild or mutant type of Apaf-1 3′ UTR was cloned into pMIR-REPORT plasmid and co-transfected with miR-27a mimics or antioligonucleotides. Dual-Luciferase Reporter Assay System was used to detect the luciferase activities. *P < 0.05. #P < 0.05. (E) Effect of miR-27a mimics and antioligonucleotides on changing the expression of Apaf-1 in HT29 CSCs and non-CSCs.