Fig. 2: Relationship between the IGF-1R gene and hsa-miR-12528 in A549 cells. | Cell Death & Disease

Fig. 2: Relationship between the IGF-1R gene and hsa-miR-12528 in A549 cells.

From: The novel hsa-miR-12528 regulates tumourigenesis and metastasis through hypo-phosphorylation of AKT cascade by targeting IGF-1R in human lung cancer

Fig. 2

The target validation of miR-12528. The pGL3-plasmid was recombined by inserting the 3′-UTR of IGF-1R. The binding sequence of miR-12528 is marked in yellow fills (top; wild type, WT), and was replaced with a mismatched sequence using site-directed mutagenesis (bottom; mutant, MT) (a). Luciferase activity of WT and MT was analysed by co-transfecting the recombinant pGL3 construction, pRL-TK and miRNA mimics, and normalised to pRL-TK; Renilla luciferase (b). The expression levels between mRNA and protein of IGF-1R gene were assessed using quantitative RT-PCR and western blot analysis and normalised to 18S rRNA or GAPDH. Western blot analysis was semi-quantified using NIH ImageJ programme (c). The signalling alteration of the IGF-1R-downstream pathway was analysed using a western blot analysis 48 h post transfection (mimic conc.; 50 nM and *; 100 nM) (d) (bars, mean ± S.E.M; *p < 0.05 and **p < 0.01)

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