Figure 5

Activation of NF-kB pathways in tubular epithelial cells with type I IFN signature. (A) RPTEC were stimulated at different time points with IFN-alpha 100 U/ml and stained for p65 (green) and pNIK (red). At basal conditions both proteins were detectable only in the cytoplasm; after five minutes of stimulation p65 and pNIK began to move from the cytoplasm to the nucleus with significant translocation after 15 and 30 minutes. (B) Specific p65 and pNIK fluorescence intensity inside the nuclei was quantified as described in Methods section. Data are averages ± SD for n = 10 cells from one field on one slide; *P <0.05 and °P <0.05 for comparison pNIK and p65 versus basal, respectively. The images and results are representative of at least three independent experiments. (C) Double staining for MXA (green), p65 (red) and merge analysis (yellow) on class IV lupus nephritis showing translocation of p65 from cytoplasm to nuclei. NF-kB, nuclear factor-kB; RPTEC, renal proximal tubular cells.