Fig. 4

The effect of Akt knockdown in WI-38 cells double-treated with GA and LPS. aWI-38 cells were transfected with a siRNA specifically targeting human Akt gene (si_Akt), and a non-specific control siRNA (si_C). Then, a qRT-PCR assay was performed to compare WI-38 endogenous Akt expressions (* P < 0.05). bCell viabilities was compared among WI-38 cells under control (ctrl) condition (not transfected by siRNA, nor treated with LPS), WI-38 cells transfected with si_C or si_Akt, and siRNA-transfected WI-38 cells treated with 10mg/ml LPS for 24h (* P < 0.05; ∆ P > 0.05). cSiRNA-transfected WI-38 cells were pre-incubated with 50 nM GA for 24h and then treated with 10mg/ml LPS for another 24h (LPS + GA). A TUNEL assay was applied to identify apoptotic cells with TUNEL (Red) immunoreaction. Also, DAPI (Blue) immunoreaction was applied to identify WI-38 cell nuclei. dFor siRNA-transfected WI-38 cells which were treated with LPS + GA, the percentages of apoptotic WI-38 cells were compared (* P < 0.05). eFor siRNA-transfected WI-38 cells which were treated with LPS + GA, western blot analysis was conducted to examine IL-6 and MCP-1 protein expressions. fFor western blot data in (E), quantitative assessment on relative band intensities of IL-6 and MCP-1 were performed (* P < 0.05)