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. 2017 May 17;6(2):18. doi: 10.3390/pathogens6020018

Figure 2.

Figure 2

ETEC FliC blocks TNF-induced IκBα degradation. (A) IκBα immunoreactivity after incubating HCT-8 cells with WT and mutant ETEC supernatants (10 µg protein) and then stimulating the cells with TNF (20 ng/mL, 20 min); (B) Purified recombinant FliC resolved using 10% SDS-PAGE and Coomassie blue staining; (C) IκBα immunoreactivity after incubating HCT-8 cells with FliC-pET28a-BL21 (100 pg–10 µg) for 1.5 h followed by TNF stimulation (20 ng/mL, 20 min); (D) IκBα immunoreactivity after incubating HCT-8 cells with FliC-pET28a-ClearColi (100 pg–10 µg) for 1.5 h and then stimulating the cells with TNF (20 ng/mL, 20 min); (E) IκBα immunoreactivity after incubating HCT-8 cells with heated (100 °C, 20 min), recombinant FliC (1 µg) for 1.5 h followed by TNF stimulation (20 ng/mL, 20 min); (F) IκBα immunoreactivity after pretreating HCT-8 cells with 1.5 µg/mL hTRL5-Fc for 1 h, followed by FliC (100 ng/mL, 90 min) and TNF (20 ng/mL, 20 min).