Fig. 4

The effect of E343 mutation on inflammatory response. HeLa Cells were stimulated with SDF-1 (50 nM) for 0, 4 and 6 h, respectively. The mRNA expression level of TNF-α (a) and IL-6 (b) was quantified by qRT-PCR. Western blotting was performed for phosphorylated PI3K (c) and AKT (e). Total PI3K and AKT were used as loading control. The relative phosphorylation level was quantified using Image J software and showed in (d) for PI3K and (f) for AKT, respectively. The subcellular localization of p65 was photographed using confocal microscope (g) and relative phosphorylated p65 which localized in the nucleus were quantified using ImageJ software (h). Bars represent the mean value ± standard deviation (SD). * P < 0.05; ** P < 0.01; *** P < 0.001