Fig. 6

Fig. 6

S-protein activates NF-κB pathways in ARPE-19 cells. A Immunofluorescence staining of p65 protein in S-protein-treated or Flag-S-overexpressed ARPE-19 cells. The nuclei were stained with DAPI. The scale bar represents 20 μm. B Cytosol-Nucleus fraction assay to determine the localization of p65 in ARPE-19 cells that were treated with PBS (Con) or S-protein for 48 h. BrgI and GAPDH were used as nuclear and cytosolic markers respectively. C Immunoblot of iκb and GAPDH in ARPE-19 cells that were treated with sham, S-protein and S-protein + NAC respectively. D Densitometry quantitation of iκb vs GAPDH in C. The data represent mean ± SD (n = 3), *p < 0.05. E Immunoblot of the expression of p53, p21 and GAPDH in ARPE-19 cells that were treated with sham, S-protein and S-protein + Bay-11–7082 (NF-κB inhibitor) for 24 h. F Densitometry quantitation of P53 and p21 in E. The P53 and P21 protein level was normalized to GAPDH. The data represent mean ± SD (n = 3), *p < 0.05. G qPCR measuring the expression of p53, p14arf, IL-1β, IL-6, TGF-β1 and iCAM in ARPE-19 cells treated in E. The data represent mean ± SD (n = 3), **p < 0.01, ***p < 0.001