Fig. 4

LPS activated NF-κB through the non-canonical pathway. (a) Significantly changing proteins were compared with known NFKB1 p50 and NFKB2 p52 targets [39]. Western blotting analysis for NFKB1 p105/p50 (b) and NFKB2 p100/p52 (c) following a 1-h exposure to TNF, IL-1β, and LPS. (d) Western blotting analysis for NFKB2 p100/p52 following a 24-h exposure and plot for p100/p52 ratio and abundances of p100 and p52. Protein abundances were normalized using the intensity of GAPDH. Blots are representative of three biological replicates, and data are represented as means ± SD. One-way ANOVA with Dunnet’s post hoc test was used to test for significance in each group compared to the control (*P < 0.05, **P < 0.01, and ***P < 0.005). (e) Confocal images of cells immuno-labeled for RelA following a 1-h or 8-h exposure to TNF, IL-1β, and LPS. Nuclei were counterstained with DAPI. The images are representative of two biological replicates. Scale bar, 50 μm