FIG. 3.

Results of AKT2 gene knockdown in MC3T3-E1 cells. (A) Fluorescence photograph of MC3T3-E1 cells transfected with AKT2 RNAi plasmids. (a) AKT2-1; (b) AKT2-2; (c) AKT2-3; (d) cells transfected with scrambled RNAi; (e) normal MC3T3-E1 cells; (f) normal MC3T3-E1 cells detected under optical microscope. (B) RT-PCR analysis of AKT2 mRNA expression in MC3T3-E1 cells transfected with NS, scrambled plasmids; WT, untransfected cells; AKT2 RNAi plasmids AKT2-1, AKT2-2, and AKT2-3 (^ap<0.05 vs. NS). (C) Western blot analysis of p-AKT, AKT, p-AKT2, and AKT2 protein expression in cells. (a) AKT and p-AKT protein expression in MC3T3-E1 cells treated with different concentrations of LY294002 with or without BRS; (b) AKT2 and p-AKT2 expression in AKT2 knockdown MC3T3-E1 cells; NS: scrambled plasmids; WT: untransfected cells. (D) The relative intensity of p-AKT, AKT, p-AKT2, and AKT2 protein in cells. (a) The relative intensity of p-AKT and AKT protein in the N0 and S0 group (*p<0.05 vs. N0). (b) The relative intensity of the p-AKT and AKT protein in the S0, S1, S2, and S3 group (^ap<0.05 vs. S0, ^bp<0.05 vs. S1, ^cp<0.05 vs. S2). (c) The relative intensity of the p-AKT and AKT protein in the N0, N1, N2, and N3 group (^ap<0.05 vs. N0, ^bp<0.05 vs. N1, ^c p<0.05 vs. N2). (d) The relative intensity of the p-AKT2 and AKT2 protein in the WT, NS, and AKT2-1 group (^#p<0.05 vs. WT). Color images available online at www.liebertpub.com/tea