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A. Early activation of potassium channels in patients inoculated with E. coli 83972 fim ( n = 47). GSEA, 3-hour samples from P I–P IV. B. Kinetics of ion channel expression in patients inoculated with E. coli 83972 fim , showing a rapid Ca 2+ and K + channels response at 3 hours and a sustained K + channels response at 24- and 48 hours. C-E. Increased expression of K + channels (TWIK, TRAAK and KCNJ11 but not KCNJ2, C ) and cation channels <t>(TRPC1</t> and TRPV6, D and E ) in bladder epithelial cells infected with E. coli 83972 or E. coli 83972 fim (10 5 cfu/ml, 4 hours). Confocal imaging ( C,D ) and Western blot analysis ( E ). Scale bars = 20 μm. F. The increase in TRPC1 and TRPV6 expression was effectively blocked by addition of the soluble FimH antagonist α-D-methyl-mannopyranoside (α-D-man., 2.5%). G,H. Purified FimCH protein (1.25–5 μg/ml, 4 hours) increased TRPC1 expression in a dose dependent manner. Confocal imaging ( G ) and Western blot analysis ( H ). Scale bars = 20 μm. I. Activation of K + , Ca 2+ and Zn 2+ fluxes by purified FimCH (5 μg/ml) as determined by fluorescence spectrometry with repeated 20 second-measurements for 16-20 minutes (mean of three experiments). The responses were effectively blocked by addition of α-D-methyl-mannopyranoside (α-D-man., 2.5%). Mean + s.e.m. of three experiments, 50 cells/experiment. Two-tailed unpaired t -test compared to PBS. P < 0.05 (*), P < 0.01 (**), P < 0.001 (***).
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A. Early activation of potassium channels in patients inoculated with E. coli 83972 fim ( n = 47). GSEA, 3-hour samples from P I–P IV. B. Kinetics of ion channel expression in patients inoculated with E. coli 83972 fim , showing a rapid Ca 2+ and K + channels response at 3 hours and a sustained K + channels response at 24- and 48 hours. C-E. Increased expression of K + channels (TWIK, TRAAK and KCNJ11 but not KCNJ2, C ) and cation channels <t>(TRPC1</t> and TRPV6, D and E ) in bladder epithelial cells infected with E. coli 83972 or E. coli 83972 fim (10 5 cfu/ml, 4 hours). Confocal imaging ( C,D ) and Western blot analysis ( E ). Scale bars = 20 μm. F. The increase in TRPC1 and TRPV6 expression was effectively blocked by addition of the soluble FimH antagonist α-D-methyl-mannopyranoside (α-D-man., 2.5%). G,H. Purified FimCH protein (1.25–5 μg/ml, 4 hours) increased TRPC1 expression in a dose dependent manner. Confocal imaging ( G ) and Western blot analysis ( H ). Scale bars = 20 μm. I. Activation of K + , Ca 2+ and Zn 2+ fluxes by purified FimCH (5 μg/ml) as determined by fluorescence spectrometry with repeated 20 second-measurements for 16-20 minutes (mean of three experiments). The responses were effectively blocked by addition of α-D-methyl-mannopyranoside (α-D-man., 2.5%). Mean + s.e.m. of three experiments, 50 cells/experiment. Two-tailed unpaired t -test compared to PBS. P < 0.05 (*), P < 0.01 (**), P < 0.001 (***).
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A. Early activation of potassium channels in patients inoculated with E. coli 83972 fim ( n = 47). GSEA, 3-hour samples from P I–P IV. B. Kinetics of ion channel expression in patients inoculated with E. coli 83972 fim , showing a rapid Ca 2+ and K + channels response at 3 hours and a sustained K + channels response at 24- and 48 hours. C-E. Increased expression of K + channels (TWIK, TRAAK and KCNJ11 but not KCNJ2, C ) and cation channels <t>(TRPC1</t> and TRPV6, D and E ) in bladder epithelial cells infected with E. coli 83972 or E. coli 83972 fim (10 5 cfu/ml, 4 hours). Confocal imaging ( C,D ) and Western blot analysis ( E ). Scale bars = 20 μm. F. The increase in TRPC1 and TRPV6 expression was effectively blocked by addition of the soluble FimH antagonist α-D-methyl-mannopyranoside (α-D-man., 2.5%). G,H. Purified FimCH protein (1.25–5 μg/ml, 4 hours) increased TRPC1 expression in a dose dependent manner. Confocal imaging ( G ) and Western blot analysis ( H ). Scale bars = 20 μm. I. Activation of K + , Ca 2+ and Zn 2+ fluxes by purified FimCH (5 μg/ml) as determined by fluorescence spectrometry with repeated 20 second-measurements for 16-20 minutes (mean of three experiments). The responses were effectively blocked by addition of α-D-methyl-mannopyranoside (α-D-man., 2.5%). Mean + s.e.m. of three experiments, 50 cells/experiment. Two-tailed unpaired t -test compared to PBS. P < 0.05 (*), P < 0.01 (**), P < 0.001 (***).
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Image Search Results


A. Early activation of potassium channels in patients inoculated with E. coli 83972 fim ( n = 47). GSEA, 3-hour samples from P I–P IV. B. Kinetics of ion channel expression in patients inoculated with E. coli 83972 fim , showing a rapid Ca 2+ and K + channels response at 3 hours and a sustained K + channels response at 24- and 48 hours. C-E. Increased expression of K + channels (TWIK, TRAAK and KCNJ11 but not KCNJ2, C ) and cation channels (TRPC1 and TRPV6, D and E ) in bladder epithelial cells infected with E. coli 83972 or E. coli 83972 fim (10 5 cfu/ml, 4 hours). Confocal imaging ( C,D ) and Western blot analysis ( E ). Scale bars = 20 μm. F. The increase in TRPC1 and TRPV6 expression was effectively blocked by addition of the soluble FimH antagonist α-D-methyl-mannopyranoside (α-D-man., 2.5%). G,H. Purified FimCH protein (1.25–5 μg/ml, 4 hours) increased TRPC1 expression in a dose dependent manner. Confocal imaging ( G ) and Western blot analysis ( H ). Scale bars = 20 μm. I. Activation of K + , Ca 2+ and Zn 2+ fluxes by purified FimCH (5 μg/ml) as determined by fluorescence spectrometry with repeated 20 second-measurements for 16-20 minutes (mean of three experiments). The responses were effectively blocked by addition of α-D-methyl-mannopyranoside (α-D-man., 2.5%). Mean + s.e.m. of three experiments, 50 cells/experiment. Two-tailed unpaired t -test compared to PBS. P < 0.05 (*), P < 0.01 (**), P < 0.001 (***).

Journal: PLoS Pathogens

Article Title: Fimbriae reprogram host gene expression – Divergent effects of P and type 1 fimbriae

doi: 10.1371/journal.ppat.1007671

Figure Lengend Snippet: A. Early activation of potassium channels in patients inoculated with E. coli 83972 fim ( n = 47). GSEA, 3-hour samples from P I–P IV. B. Kinetics of ion channel expression in patients inoculated with E. coli 83972 fim , showing a rapid Ca 2+ and K + channels response at 3 hours and a sustained K + channels response at 24- and 48 hours. C-E. Increased expression of K + channels (TWIK, TRAAK and KCNJ11 but not KCNJ2, C ) and cation channels (TRPC1 and TRPV6, D and E ) in bladder epithelial cells infected with E. coli 83972 or E. coli 83972 fim (10 5 cfu/ml, 4 hours). Confocal imaging ( C,D ) and Western blot analysis ( E ). Scale bars = 20 μm. F. The increase in TRPC1 and TRPV6 expression was effectively blocked by addition of the soluble FimH antagonist α-D-methyl-mannopyranoside (α-D-man., 2.5%). G,H. Purified FimCH protein (1.25–5 μg/ml, 4 hours) increased TRPC1 expression in a dose dependent manner. Confocal imaging ( G ) and Western blot analysis ( H ). Scale bars = 20 μm. I. Activation of K + , Ca 2+ and Zn 2+ fluxes by purified FimCH (5 μg/ml) as determined by fluorescence spectrometry with repeated 20 second-measurements for 16-20 minutes (mean of three experiments). The responses were effectively blocked by addition of α-D-methyl-mannopyranoside (α-D-man., 2.5%). Mean + s.e.m. of three experiments, 50 cells/experiment. Two-tailed unpaired t -test compared to PBS. P < 0.05 (*), P < 0.01 (**), P < 0.001 (***).

Article Snippet: Primary rabbit antibodies: anti–IRF-7 antibody (1:200, ab62505, Abcam), anti-PapG pre-absorbed serum (1:1,000), anti-TWIK-1 (1:50 sc-28630, Santa Cruz biotechnologies), anti-TRAAK (1:50, sc-50413, Santa Cruz biotechnologies), anti-KCNJ2 (1:100, 3305–1, Epitomics), anti-KCNJ11 (1:100 APC-202, Alomone Labs), anti-TRPC1 (1:100, ACC-010, Alomone Labs), anti-TRPV6 (1:250 orb158655, Biorbyt) and secondary goat anti-rabbit Alexa Fluor 488–conjugated antibody (1:200, A-11034, Thermo Fisher Scientific) were used.

Techniques: Activation Assay, Expressing, Infection, Imaging, Western Blot, Purification, Fluorescence, Two Tailed Test