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. 2017 Sep 8;7:11039. doi: 10.1038/s41598-017-10445-4

Figure 1.

Figure 1

VK2 E6/E7 and RT4 DEFB4 and BD2 responses to UPEC and flagellin challenges. End point PCR panels of host defence peptide gene expression in VK2 E6/E7 and RT4 cells following 24 hr challenge with PBS (control), flagellin (250 ng/ml) or UPEC (105) (A). Full length gels are shown in Supplementary File 1 (S1A–D). DEFB4 transcript expression in VK2 E6/E7 and RT4 cells following challenges with flagellin (250 ng/ml) and heat-killed flagellated (NCTC10418) or non-flagellated (NU14) E. coli (105) (N = 3, n = 9) (B). BD2 concentrations measured in VK2 E6/E7 and RT4 cell media following 24 hour challenges with flagellin (250 ng/ml) and heat-killed CFT073 (105) (N = 3, n = 6) (C). BD2 peptide concentrations of primary ureteric urothelial cells challenged with E.coli flagellin (250 ng/ml) for 24 hours (N = 3, n = 6) (D). DEFB4 transcript expression, presented as relative expression, of primary vaginal epithelial cells challenged with E.coli flagellin (50 ng/ml) for 24 hours (N = 6) (E). E.coli swarming motility on motility agar media; BD2 peptide concentrations measured in VK2 E6/E7 and RT4 cell media following 24 hour challenges with flagellin (250 ng/ml) and heat-killed bacteria (105) (N = 3, n = 6) (F). DEFB4 transcript expression of VK2 E6/E7 cells transfected with either negative siRNA or TLR5 siRNA and challenged for 24 hours with flagellin (50ng/ml) (N = 3) (G). BD2 peptide concentrations of VK2 E6/E7 cells incubated with TLR5 blocking antibody (1–5 ug/ml) or IgG (5 ug/ml) and challenged 24 hours with flagellin (250 ng/ml) (N = 3) (H). Fold change in NFĸB-luciferase activity in RT4 cells challenged with 250 ng/ml flagellin (FLG) for 2–8 hours, peptidoglycan (PG) (10 μg/ml) for up to 16 hours and Lipopolysaccharide (LPS) (10 μg/ml) for up to 16 hours (N = 3, n = 6 (FLG), n = 9 (PG & LPS)) (I). All data presented as mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001.