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. 2019 May 27;9:7903. doi: 10.1038/s41598-019-44263-7

Figure 5.

Figure 5

Flagellin increases virus entry through NF-kB and signalling. (a) A549 cells were treated with STm FliC (1 μg/mL) for 1 h and media replaced for various time periods prior to inoculating with VSV-Gpp for 1 h and luciferase expression assessed after 48 h. (b) A549 cells were transfected with an NF-ΚB reporter plasmid (pConA-luciferase) for 24 h, inoculated with STm (MOI 10), STm CM or STm FliC (0.3 μg/mL) for 1 h prior to chloramphenicol (34 μg/mL) addition and NF-ΚB activity measured after 24 h. NF-ΚB reporter transfected cells were treated with MLN4924 for 15 mins and inoculated with STm (MOI 10) for 1 h prior to chloramphenicol (34 μg/mL) addition and NF-ΚB activity measured. (c) A549 cells were treated with MLN4924 (1 uM) for 15 mins and inoculated with STm (MOI 10), STm CM or STm FliC (0.3 μg/mL) for 1 h prior to inoculating with VSV-Gpp and chloramphenicol (34 μg/mL) and VSV-Gpp entry or interleukin 8 (IL-8) expression. (d) A549 cells were transfected with control siRNA or RelA siRNA for 24 h prior to lysing for western blotting. Transfected cells were exposed to STm flagellin (0.3 μg/mL) for 1 h prior to inoculating with VSV-Gpp and entry determined. Bars represent mean ± S.D. for n = 3. Statistical comparison by unpaired t test where: n.s. p > 0.05; *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001 and ****p ≤ 0.0001. All data sets are representative of at least two independent experiments.