Journal: Scientific reports

Article Title: Targeting Deficiencies in the TLR5 Mediated Vaginal Response to Treat Female Recurrent Urinary Tract Infection.

doi: 10.1038/s41598-017-10445-4

Figure Lengend Snippet: 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

Article Snippet: VK2 E6/E7 cells (ATCC CRL-2616)54 were maintained without antibiotics in keratinocyte serum-free medium (GIBCO, Paisley, UK) containing 0.4 mM calcium with 0.1 ng/ml human recombinant Epidermal Growth Factor (EGF) and 0.05 mg/ml bovine pituitary extract supplements.

Techniques: Gene Expression, Control, Expressing, Bacteria, Transfection

Journal: Scientific reports

Article Title: Targeting Deficiencies in the TLR5 Mediated Vaginal Response to Treat Female Recurrent Urinary Tract Infection.

doi: 10.1038/s41598-017-10445-4

Figure Lengend Snippet: Figure 5. Effects of estrogen treatment on BD2 responses in vitro and rUTI patients. VK2 E6/E7 cell DEFB4 mRNA expression following estrogen (4 nM) seven day pretreatment and 24 h challenge with either cyclodextrin (15 nM), flagellin (50 ng/ml), or flagellin (50 ng/ml) and estrogen (4 nM) N = 3 (A). BD2 peptide concentrations measured in the VK2 E6/E7 cell media following estrogen (4 nM) seven day pretreatment and either 24 h or 48 h challenge with either cyclodextrin (15 nM), estrogen (4 nM), flagellin (50 ng/ml) or flagellin and estrogen (N = 3) (B). Vaginal douche BD2 peptide concentrations of rUTI patients before and after 6–8 weeks of treatment with antibiotic prophylaxis and/or advice (N = 12) (C), or 6 to 8 weeks treatment with vaginal estrogen (Vagifem, 10 mcg, twice weekly) (N = 9) (D). Staining of vaginal tissue from estrogen treated ( + E) and control subjects for immunoreactive BD2 (E). Vaginal douche BD2 peptide concentrations of rUTI TLR5 SNP patients before and after treatment (6 to 8 weeks) with vaginal estrogen (N = 3) or antibiotic

Article Snippet: VK2 E6/E7 cells (ATCC CRL-2616)54 were maintained without antibiotics in keratinocyte serum-free medium (GIBCO, Paisley, UK) containing 0.4 mM calcium with 0.1 ng/ml human recombinant Epidermal Growth Factor (EGF) and 0.05 mg/ml bovine pituitary extract supplements.

Techniques: In Vitro, Expressing, Staining, Control