RLN2 Is a Positive Regulator of AKT-2-Induced Gene Expression Required for Osteosarcoma Cells Invasion and Chemoresistance

Figure 3

Effect of RLN2 inhibition decreased AKT/NF-κB signaling pathway in MG-63 cells. MG-63 cells were transfected with RLN2 siRNA2 for 48 hs and then treated with myr-AKT or TNF-α at different time points. (a) The protein of p-AKT (Ser473), p-ERK1/2, NF-κB (p65), MMP-9, VEGF, and bcl-2 was measured using western blot analysis. (b) Nuclear extracts were prepared and subjected to analysis for NF-κB DNA-binding activity as measured by EMSA. Retinoblastoma protein level served as nuclear protein loading control. (c) Zymographic analysis of MMP-9 gelatinolytic activity, which was quantified by densitometry and graphed in OD units (mean ± SEM) ().

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