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. 2017 Mar 30;8(26):41921–41931. doi: 10.18632/oncotarget.16707

Figure 5. Effect of miR-106b inhibitors on TRAIL-dependent apoptotic pathway.

Figure 5

(A) After treatment with TRAIL (2 ng/ml), anti-miR-106b (50 pmol/ml) and DR4 siRNA (50 pmol/ml), western blot analysis was performed to evaluate the effect of anti-miR-106b on TRAIL-dependent activation of caspase-8. (B) After treatment with TRAIL (2 ng/ml), anti-miR-106b (50 pmol/ml) and DR4 siRNA (50 pmol/ml), mitochondria were seperated from Huh7 and HepG2 cells, expression level of tBid was evaluated by western blot analysis. Cox IV was detected as internal control of mitochondrial proteins. (C) After treatment with TRAIL (2 ng/ml), anti-miR-106b (50 pmol/ml) and DR4 siRNA (50 pmol/ml), mitochondrial membrane potential (MMP) of Huh7 and HepG2 cells was detected by flow cytometry by using JC-1 staining. (D) After treatment with TRAIL (2 ng/ml), anti-miR-106b (50 pmol/ml) and DR4 siRNA (50 pmol/ml), cleavage of caspase-9, caspase-3 and PARP in TRAIL signaling pathway was detected by western blot analysis. (E) After treatment with TRAIL (2 ng/ml), anti-miR-106b (50 pmol/ml) and DR4 siRNA (50 pmol/ml), cell apoptosis of Huh7 and HepG2 was measured after they were treated with TRAIL and RNA oligoribonucleotides.