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Figure 3 | Molecular Cancer

Figure 3

From: Comprehensive analysis of microRNA-regulated protein interaction network reveals the tumor suppressive role of microRNA-149 in human hepatocellular carcinoma via targeting AKT-mTOR pathway

Figure 3

MiRNA-149 directly targets AKT1 in HCC cells. (A) QRT-PCR analysis showing relative expression of miR-149 in HepG2 cells transfected with miR-149 mimics, miR-149 mimic control (negative control, NC), and blank control culture medium (mock). (B, C and F) Relative expression of AKT1, p-AKT1, mTOR and p-mTOR proteins in HepG2 cells transfected with miR-149 mimics, miR-149 mimic control (negative control, NC), and blank control culture medium (mock) detected by Western blot analysis. GAPDH was used as an internal loading control. (D) RNA sequence alignment showing the 3′-UTR of AKT1 mRNA contains a complementary site for the seed region of miR-149. AKT1mut is amutant with substitutions in the complementary region as a negative control. (E) Luciferase report assay was performed to verify whether AKT1 was a direct target of miR-149. The luciferase activity was detected after transfection of FLuci vector (3′-UTR-AKT1wt FLuci vector or 3′-UTR-AKT1mut FLuci vector) into the miR-149 mimic or miR-149 mimic control (negative control, NC) transfected HepG2 cells.

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