t84 colonic cancer epithelial cells Search Results


96
ATCC human t84 colon epithelial cells
Human T84 Colon Epithelial Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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96
Thermo Fisher hplc pure 4c
Hplc Pure 4c, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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90
DS Pharma Biomedical human epithelial colonic carcinoma cell line t84
Human Epithelial Colonic Carcinoma Cell Line T84, supplied by DS Pharma Biomedical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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99
ATCC epithelial cells lines
<t> Epithelial cells </t> used in this study
Epithelial Cells Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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90
Millipore t84 cells
<t> Epithelial cells </t> used in this study
T84 Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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99
ATCC cc lines
<t> Epithelial cells </t> used in this study
Cc Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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90
Koehler Instrument t84 intestinal epithelial cells
<t> Epithelial cells </t> used in this study
T84 Intestinal Epithelial Cells, supplied by Koehler Instrument, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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90
European Collection of Authenticated Cell Cultures intestinal epithelial cell line t84
A: <t>T84</t> cells were cultured in a Transwell system and then infected with several types of A . sobria strains. After 3 hr of infection (MOI = 5), the reduction in TER was measured. The TER value at 0 hr of infection was taken as 100%. NT: The TER value was measured without bacterial infection. The experiments were performed in triplicate. The data are mean ± SD (error bars). *p<0.01. B: The ability of the A . sobria strains to translocate across the intestinal <t>epithelial</t> cells (T84 cells) at 6 hr after infection (MOI = 5) was assessed using quantitative cultures of medium obtained from the lower chambers. NT: The experiment was done without bacterial infection. ND: The bacterial translocation could not be detected in this experimental condition. The experiments were performed in triplicate. The data are mean ± SD (error bars). C: The proteolytic activity in the culture supernatant of each A . sobria strain was measured as described in the text. The experiments were performed in triplicate. The data are mean ± SD (error bars). D: The presence of ASP in the culture supernatant of each strain was immunologically detected by a western blotting analysis as described in the text.
Intestinal Epithelial Cell Line T84, supplied by European Collection of Authenticated Cell Cultures, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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90
Flow Laboratories t84 (derived from human colon carcinoma)
A: <t>T84</t> cells were cultured in a Transwell system and then infected with several types of A . sobria strains. After 3 hr of infection (MOI = 5), the reduction in TER was measured. The TER value at 0 hr of infection was taken as 100%. NT: The TER value was measured without bacterial infection. The experiments were performed in triplicate. The data are mean ± SD (error bars). *p<0.01. B: The ability of the A . sobria strains to translocate across the intestinal <t>epithelial</t> cells (T84 cells) at 6 hr after infection (MOI = 5) was assessed using quantitative cultures of medium obtained from the lower chambers. NT: The experiment was done without bacterial infection. ND: The bacterial translocation could not be detected in this experimental condition. The experiments were performed in triplicate. The data are mean ± SD (error bars). C: The proteolytic activity in the culture supernatant of each A . sobria strain was measured as described in the text. The experiments were performed in triplicate. The data are mean ± SD (error bars). D: The presence of ASP in the culture supernatant of each strain was immunologically detected by a western blotting analysis as described in the text.
T84 (Derived From Human Colon Carcinoma), supplied by Flow Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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99
ATCC ileocecal carcinoma dmem f 12 d sheila crowe
A: <t>T84</t> cells were cultured in a Transwell system and then infected with several types of A . sobria strains. After 3 hr of infection (MOI = 5), the reduction in TER was measured. The TER value at 0 hr of infection was taken as 100%. NT: The TER value was measured without bacterial infection. The experiments were performed in triplicate. The data are mean ± SD (error bars). *p<0.01. B: The ability of the A . sobria strains to translocate across the intestinal <t>epithelial</t> cells (T84 cells) at 6 hr after infection (MOI = 5) was assessed using quantitative cultures of medium obtained from the lower chambers. NT: The experiment was done without bacterial infection. ND: The bacterial translocation could not be detected in this experimental condition. The experiments were performed in triplicate. The data are mean ± SD (error bars). C: The proteolytic activity in the culture supernatant of each A . sobria strain was measured as described in the text. The experiments were performed in triplicate. The data are mean ± SD (error bars). D: The presence of ASP in the culture supernatant of each strain was immunologically detected by a western blotting analysis as described in the text.
Ileocecal Carcinoma Dmem F 12 D Sheila Crowe, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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90
European Collection of Authenticated Cell Cultures t84 cells (ecacc 88021101)
qRT-PCR showing expression of human (A) CHOP , (B) spliced XBP1 [ XBP1(s) ], (C) ATF6 and (D) BiP in <t>T84</t> cells infected with C. jejuni 11168H, 81-176 or 488 wild-type strains for 6 or 24 h at 37°C in a 5% CO 2 incubator (MOI of 200:1). T84 cells were treated with thapsigargin as a positive control. GAPDH was used as an internal control. One sample t -test was performed. Asterisks denote a statistically significant difference (** = p < 0.01; *** = p < 0.001; **** = p < 0.0001).
T84 Cells (Ecacc 88021101), supplied by European Collection of Authenticated Cell Cultures, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


 Epithelial cells  used in this study

Journal:

Article Title: Airway epithelial cells synthesize the lipid mediator 5-oxo-ETE in response to oxidative stress

doi: 10.1016/j.freeradbiomed.2006.12.006

Figure Lengend Snippet: Epithelial cells used in this study

Article Snippet: Epithelial cells lines (A549, BEAS-2B, T84, and HEp-2) were obtained from the American Type Culture Collection ( ) and cultured in 10% FBS in DMEM/F12 medium containing L-glutamine (2 mM), sodium bicarbonate (2.5 g/L), penicillin (100U/ml), and streptomycin (100 μg/ml).

Techniques:

Epithelial cells were cultured to confluence in 6-well plates. Cells were washed twice with PBS+ and then preincubated for 6 min with 100 μM PMS. The cells were then incubated for an additional 20 min with 4 μM 5-HETE. Cells in adjoining wells were detached with trypsin and counted. Values are means ± SE (n = 4, except for T84 and HEp-2 cells (n=2))..

Journal:

Article Title: Airway epithelial cells synthesize the lipid mediator 5-oxo-ETE in response to oxidative stress

doi: 10.1016/j.freeradbiomed.2006.12.006

Figure Lengend Snippet: Epithelial cells were cultured to confluence in 6-well plates. Cells were washed twice with PBS+ and then preincubated for 6 min with 100 μM PMS. The cells were then incubated for an additional 20 min with 4 μM 5-HETE. Cells in adjoining wells were detached with trypsin and counted. Values are means ± SE (n = 4, except for T84 and HEp-2 cells (n=2))..

Article Snippet: Epithelial cells lines (A549, BEAS-2B, T84, and HEp-2) were obtained from the American Type Culture Collection ( ) and cultured in 10% FBS in DMEM/F12 medium containing L-glutamine (2 mM), sodium bicarbonate (2.5 g/L), penicillin (100U/ml), and streptomycin (100 μg/ml).

Techniques: Cell Culture, Incubation

Scheme showing the regulation of 5-oxo-ETE synthesis in airway epithelial cells. Metabolism of H2O2 and tBuOOH by glutathione peroxidase (GPerox) results in the formation of GSSG, which is reduced back to GSH by glutathione reductase (GRed), generating NADP+, which serves as a cofactor for 5-HEDH. This process can be blocked by NEM, which alkylates GSH, and mimicked by diamide (DA), which oxidizes GSH, and PMS, which oxidizes NADPH. In A549 cells the action of H2O2 is limited by metabolism by catalase, which can be blocked by azide.

Journal:

Article Title: Airway epithelial cells synthesize the lipid mediator 5-oxo-ETE in response to oxidative stress

doi: 10.1016/j.freeradbiomed.2006.12.006

Figure Lengend Snippet: Scheme showing the regulation of 5-oxo-ETE synthesis in airway epithelial cells. Metabolism of H2O2 and tBuOOH by glutathione peroxidase (GPerox) results in the formation of GSSG, which is reduced back to GSH by glutathione reductase (GRed), generating NADP+, which serves as a cofactor for 5-HEDH. This process can be blocked by NEM, which alkylates GSH, and mimicked by diamide (DA), which oxidizes GSH, and PMS, which oxidizes NADPH. In A549 cells the action of H2O2 is limited by metabolism by catalase, which can be blocked by azide.

Article Snippet: Epithelial cells lines (A549, BEAS-2B, T84, and HEp-2) were obtained from the American Type Culture Collection ( ) and cultured in 10% FBS in DMEM/F12 medium containing L-glutamine (2 mM), sodium bicarbonate (2.5 g/L), penicillin (100U/ml), and streptomycin (100 μg/ml).

Techniques:

A: T84 cells were cultured in a Transwell system and then infected with several types of A . sobria strains. After 3 hr of infection (MOI = 5), the reduction in TER was measured. The TER value at 0 hr of infection was taken as 100%. NT: The TER value was measured without bacterial infection. The experiments were performed in triplicate. The data are mean ± SD (error bars). *p<0.01. B: The ability of the A . sobria strains to translocate across the intestinal epithelial cells (T84 cells) at 6 hr after infection (MOI = 5) was assessed using quantitative cultures of medium obtained from the lower chambers. NT: The experiment was done without bacterial infection. ND: The bacterial translocation could not be detected in this experimental condition. The experiments were performed in triplicate. The data are mean ± SD (error bars). C: The proteolytic activity in the culture supernatant of each A . sobria strain was measured as described in the text. The experiments were performed in triplicate. The data are mean ± SD (error bars). D: The presence of ASP in the culture supernatant of each strain was immunologically detected by a western blotting analysis as described in the text.

Journal: PLoS ONE

Article Title: Aeromonas sobria serine protease decreases epithelial barrier function in T84 cells and accelerates bacterial translocation across the T84 monolayer in vitro

doi: 10.1371/journal.pone.0221344

Figure Lengend Snippet: A: T84 cells were cultured in a Transwell system and then infected with several types of A . sobria strains. After 3 hr of infection (MOI = 5), the reduction in TER was measured. The TER value at 0 hr of infection was taken as 100%. NT: The TER value was measured without bacterial infection. The experiments were performed in triplicate. The data are mean ± SD (error bars). *p<0.01. B: The ability of the A . sobria strains to translocate across the intestinal epithelial cells (T84 cells) at 6 hr after infection (MOI = 5) was assessed using quantitative cultures of medium obtained from the lower chambers. NT: The experiment was done without bacterial infection. ND: The bacterial translocation could not be detected in this experimental condition. The experiments were performed in triplicate. The data are mean ± SD (error bars). C: The proteolytic activity in the culture supernatant of each A . sobria strain was measured as described in the text. The experiments were performed in triplicate. The data are mean ± SD (error bars). D: The presence of ASP in the culture supernatant of each strain was immunologically detected by a western blotting analysis as described in the text.

Article Snippet: The intestinal epithelial cell line T84 was obtained from the European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Cell Culture, Infection, Translocation Assay, Activity Assay, Western Blot

A: The asp gene of A . sobria 288 strain was knocked out. The immunological analysis using western blotting revealed that the asp -knocked-out strain (#288 ΔASP) did not produce ASP and the complemented strain (#288 ΔASP::ASP) produced ASP again. B: The proteolytic activity in the culture supernatant of each strain. The experiments were performed in triplicate. The data are mean ± SD (error bars). C: T84 cells were cultured in a Transwell system and then infected with the wild-type A . sobria strain (#288), the asp -knocked-out strain (#288 ΔASP), or the complemented strain (#288 ΔASP::ASP). After 6 hr of infection (MOI = 5), the ability of these A . sobria strains to translocate across the T84 cell monolayer was assessed in the same way as that described in the legend. The experiments were performed in triplicate. The data are mean ± SD (error bars). *p<0.01, **p<0.05.

Journal: PLoS ONE

Article Title: Aeromonas sobria serine protease decreases epithelial barrier function in T84 cells and accelerates bacterial translocation across the T84 monolayer in vitro

doi: 10.1371/journal.pone.0221344

Figure Lengend Snippet: A: The asp gene of A . sobria 288 strain was knocked out. The immunological analysis using western blotting revealed that the asp -knocked-out strain (#288 ΔASP) did not produce ASP and the complemented strain (#288 ΔASP::ASP) produced ASP again. B: The proteolytic activity in the culture supernatant of each strain. The experiments were performed in triplicate. The data are mean ± SD (error bars). C: T84 cells were cultured in a Transwell system and then infected with the wild-type A . sobria strain (#288), the asp -knocked-out strain (#288 ΔASP), or the complemented strain (#288 ΔASP::ASP). After 6 hr of infection (MOI = 5), the ability of these A . sobria strains to translocate across the T84 cell monolayer was assessed in the same way as that described in the legend. The experiments were performed in triplicate. The data are mean ± SD (error bars). *p<0.01, **p<0.05.

Article Snippet: The intestinal epithelial cell line T84 was obtained from the European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Western Blot, Produced, Activity Assay, Cell Culture, Infection

A: T84 cells were cultured in a Transwell system, and the TER value was measured in the presence of various concentrations (nM) of ASP or absence (NT) of ASP. We also examined the effect of the serine protease inhibitor PMSF on the action of ASP. B: The passive diffusion of FITC-labeled dextran molecules across the T84 monolayer (from the apical side to the basolateral side) treated with ASP was measured. All experiments were performed in triplicate. The data are mean ± SD (error bars).

Journal: PLoS ONE

Article Title: Aeromonas sobria serine protease decreases epithelial barrier function in T84 cells and accelerates bacterial translocation across the T84 monolayer in vitro

doi: 10.1371/journal.pone.0221344

Figure Lengend Snippet: A: T84 cells were cultured in a Transwell system, and the TER value was measured in the presence of various concentrations (nM) of ASP or absence (NT) of ASP. We also examined the effect of the serine protease inhibitor PMSF on the action of ASP. B: The passive diffusion of FITC-labeled dextran molecules across the T84 monolayer (from the apical side to the basolateral side) treated with ASP was measured. All experiments were performed in triplicate. The data are mean ± SD (error bars).

Article Snippet: The intestinal epithelial cell line T84 was obtained from the European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Cell Culture, Protease Inhibitor, Diffusion-based Assay, Labeling

T84 cells were treated with (nM) or without (NT) various concentrations of ASP before the extraction. After the extraction, we detected the proteins constituting the junctional complexes by using a specific antibody against each protein shown in the figure. The results of quantitative analysis of the amount of blotted protein are also shown below the image of western blotting. These experiments were performed in triplicate. The data are mean ± SD (error bars).

Journal: PLoS ONE

Article Title: Aeromonas sobria serine protease decreases epithelial barrier function in T84 cells and accelerates bacterial translocation across the T84 monolayer in vitro

doi: 10.1371/journal.pone.0221344

Figure Lengend Snippet: T84 cells were treated with (nM) or without (NT) various concentrations of ASP before the extraction. After the extraction, we detected the proteins constituting the junctional complexes by using a specific antibody against each protein shown in the figure. The results of quantitative analysis of the amount of blotted protein are also shown below the image of western blotting. These experiments were performed in triplicate. The data are mean ± SD (error bars).

Article Snippet: The intestinal epithelial cell line T84 was obtained from the European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Extraction, Western Blot

T84 cells were treated with or without (NT) 500 nM ASP for 6 hr. The cells were reacted with a specific antibody against nectin-1, nectin-2, and E-cadherin and visualized using the secondary antibody conjugated with a fluorescent substance, Cy5 or FITC. Nuclei were stained with PI. The merged images are shown in each panel. Merge 1: nectin-2 and E-cadherin, Merge 2: afadin and E-cadherin, and Merge 3: nectin-1 and E-cadherin. Z: Z-stack showing entire sample volume image was also shown.

Journal: PLoS ONE

Article Title: Aeromonas sobria serine protease decreases epithelial barrier function in T84 cells and accelerates bacterial translocation across the T84 monolayer in vitro

doi: 10.1371/journal.pone.0221344

Figure Lengend Snippet: T84 cells were treated with or without (NT) 500 nM ASP for 6 hr. The cells were reacted with a specific antibody against nectin-1, nectin-2, and E-cadherin and visualized using the secondary antibody conjugated with a fluorescent substance, Cy5 or FITC. Nuclei were stained with PI. The merged images are shown in each panel. Merge 1: nectin-2 and E-cadherin, Merge 2: afadin and E-cadherin, and Merge 3: nectin-1 and E-cadherin. Z: Z-stack showing entire sample volume image was also shown.

Article Snippet: The intestinal epithelial cell line T84 was obtained from the European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Staining

A: T84 cells were infected (MOI = 1) with or without (NT) A . sobria strain 120, 123 or 288. Bacterial internalization was confirmed using the Aeromonas -specific probe FITC-AER66 as described in Materials and Methods ( fluorescence image ). The figure shown as ‘Merge’ is a superposition of the fluorescence image ( left side ) and the blight image ( center ). B: T84 cells were infected (MOI = 1) with A . sobria strain 120, 123 or 288. The number of bacteria survived from gentamicin protection assay was determined and indicated as colony forming unit (CFU). The experiments were performed in triplicate and the data are means ± SD (error bars).

Journal: PLoS ONE

Article Title: Aeromonas sobria serine protease decreases epithelial barrier function in T84 cells and accelerates bacterial translocation across the T84 monolayer in vitro

doi: 10.1371/journal.pone.0221344

Figure Lengend Snippet: A: T84 cells were infected (MOI = 1) with or without (NT) A . sobria strain 120, 123 or 288. Bacterial internalization was confirmed using the Aeromonas -specific probe FITC-AER66 as described in Materials and Methods ( fluorescence image ). The figure shown as ‘Merge’ is a superposition of the fluorescence image ( left side ) and the blight image ( center ). B: T84 cells were infected (MOI = 1) with A . sobria strain 120, 123 or 288. The number of bacteria survived from gentamicin protection assay was determined and indicated as colony forming unit (CFU). The experiments were performed in triplicate and the data are means ± SD (error bars).

Article Snippet: The intestinal epithelial cell line T84 was obtained from the European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Infection, Fluorescence, Bacteria

qRT-PCR showing expression of human (A) CHOP , (B) spliced XBP1 [ XBP1(s) ], (C) ATF6 and (D) BiP in T84 cells infected with C. jejuni 11168H, 81-176 or 488 wild-type strains for 6 or 24 h at 37°C in a 5% CO 2 incubator (MOI of 200:1). T84 cells were treated with thapsigargin as a positive control. GAPDH was used as an internal control. One sample t -test was performed. Asterisks denote a statistically significant difference (** = p < 0.01; *** = p < 0.001; **** = p < 0.0001).

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: qRT-PCR showing expression of human (A) CHOP , (B) spliced XBP1 [ XBP1(s) ], (C) ATF6 and (D) BiP in T84 cells infected with C. jejuni 11168H, 81-176 or 488 wild-type strains for 6 or 24 h at 37°C in a 5% CO 2 incubator (MOI of 200:1). T84 cells were treated with thapsigargin as a positive control. GAPDH was used as an internal control. One sample t -test was performed. Asterisks denote a statistically significant difference (** = p < 0.01; *** = p < 0.001; **** = p < 0.0001).

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Quantitative RT-PCR, Expressing, Infection, Positive Control, Control

Western blotting showing protein levels of human CHOP and both spliced & unspliced forms of XBP1 [XBP1(u) & XBP1(s)] in T84 (left panels) and Caco-2 cells (right panels) infected with C. jejuni 11168H, 81-176 or 488 wild-type strains for 24 h at 37°C in a 5% CO 2 incubator (MOI of 200:1). T84 cells were treated with thapsigargin (Tg) as a positive control. GAPDH was used as an internal control.

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: Western blotting showing protein levels of human CHOP and both spliced & unspliced forms of XBP1 [XBP1(u) & XBP1(s)] in T84 (left panels) and Caco-2 cells (right panels) infected with C. jejuni 11168H, 81-176 or 488 wild-type strains for 24 h at 37°C in a 5% CO 2 incubator (MOI of 200:1). T84 cells were treated with thapsigargin (Tg) as a positive control. GAPDH was used as an internal control.

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Western Blot, Infection, Positive Control, Control

T84 or Caco-2 cells were pre-treated with 2 µM of thapsigargin for 6 h and infected with C. jejuni 11168H, 81-176 or 488 wild-type strains for 3 h at 37°C in a 5% CO 2 atmosphere (MOI of 200:1). For interaction assays, (A) T84 or (B) Caco-2 cells were washed with PBS, lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. For invasion assays, after infection with C. jejuni , (C) T84 or (D) Caco-2 cells were incubated with gentamicin (150 µg/ml) for 2 h to kill extracellular bacteria, then lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. For intracellular survival assays, the 2 h gentamicin treatment as for invasion assays was followed by further incubation with gentamicin (10 µg/ml) for 18 h, then (E) T84 or (F) Caco-2 cells were lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. One sample t -test was performed. Asterisks denote a statistically significant difference (** = p < 0.01; *** = p < 0.001).

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: T84 or Caco-2 cells were pre-treated with 2 µM of thapsigargin for 6 h and infected with C. jejuni 11168H, 81-176 or 488 wild-type strains for 3 h at 37°C in a 5% CO 2 atmosphere (MOI of 200:1). For interaction assays, (A) T84 or (B) Caco-2 cells were washed with PBS, lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. For invasion assays, after infection with C. jejuni , (C) T84 or (D) Caco-2 cells were incubated with gentamicin (150 µg/ml) for 2 h to kill extracellular bacteria, then lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. For intracellular survival assays, the 2 h gentamicin treatment as for invasion assays was followed by further incubation with gentamicin (10 µg/ml) for 18 h, then (E) T84 or (F) Caco-2 cells were lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. One sample t -test was performed. Asterisks denote a statistically significant difference (** = p < 0.01; *** = p < 0.001).

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Infection, Incubation, Bacteria

(A) T84 or (B) Caco-2 cells were pre-treated with 3 µM of KIRA6, 100 µM of STF-083010 or 3 µM of GSK2656157 for 4 h at 37°C in a 5% CO 2 incubator. T84 and Caco-2 cells were then infected with C. jejuni 11168H, 81-176 or 488 wild-type strains for 24 h at 37°C in a 5% CO 2 incubator (MOI of 200:1). Human IECs were further treated with 3 µM of KIRA6 and 3 µM of GSK2656157 during C. jejuni infection. T84 and Caco-2 cells were washed with PBS and incubated with gentamicin (150 µg/ml) for 2 h to kill extracellular bacteria and then lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. One sample t -test was performed. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01).

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: (A) T84 or (B) Caco-2 cells were pre-treated with 3 µM of KIRA6, 100 µM of STF-083010 or 3 µM of GSK2656157 for 4 h at 37°C in a 5% CO 2 incubator. T84 and Caco-2 cells were then infected with C. jejuni 11168H, 81-176 or 488 wild-type strains for 24 h at 37°C in a 5% CO 2 incubator (MOI of 200:1). Human IECs were further treated with 3 µM of KIRA6 and 3 µM of GSK2656157 during C. jejuni infection. T84 and Caco-2 cells were washed with PBS and incubated with gentamicin (150 µg/ml) for 2 h to kill extracellular bacteria and then lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. One sample t -test was performed. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01).

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Infection, Incubation, Bacteria

qRT-PCR showing expression of human (A, C) CHOP and (B, D) spliced XBP1 [ XBP1(s) ] in T84 or Caco-2 cells infected with either C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI of 200:1) or treated with 2 µM of thapsigargin for 24 h at 37°C in a 5% CO 2 atmosphere. Prior to C. jejuni infection or thapsigargin treatment, T84 and Caco-2 cells were treated with 3 µM of KIRA6, 100 µM of STF-083010 or 3 µM of GSK2656157 for 4 h at 37°C in a 5% CO 2 incubator. T84 and Caco-2 cells were further treated with 3 µM of KIRA6 and 3 µM of GSK2656157 during C. jejuni infection or thapsigargin treatment. GAPDH was used as an internal control. Three biological and three technical replicates were performed for each experiment. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01; *** = p < 0.001; **** = p < 0.0001).

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: qRT-PCR showing expression of human (A, C) CHOP and (B, D) spliced XBP1 [ XBP1(s) ] in T84 or Caco-2 cells infected with either C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI of 200:1) or treated with 2 µM of thapsigargin for 24 h at 37°C in a 5% CO 2 atmosphere. Prior to C. jejuni infection or thapsigargin treatment, T84 and Caco-2 cells were treated with 3 µM of KIRA6, 100 µM of STF-083010 or 3 µM of GSK2656157 for 4 h at 37°C in a 5% CO 2 incubator. T84 and Caco-2 cells were further treated with 3 µM of KIRA6 and 3 µM of GSK2656157 during C. jejuni infection or thapsigargin treatment. GAPDH was used as an internal control. Three biological and three technical replicates were performed for each experiment. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01; *** = p < 0.001; **** = p < 0.0001).

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Quantitative RT-PCR, Expressing, Infection, Control

Western blotting showing protein levels of both spliced & unspliced forms of XBP1 [XBP1(u) & XBP1(s)] and CHOP in T84 cells infected with C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI of 200:1) or treated with 2 µM of thapsigargin (Tg) for 24 h at 37°C in a 5% CO 2 incubator. Prior to C. jejuni infection or thapsigargin treatment, T84 and Caco-2 cells were pre-treated with 3 µM of KIRA6, 100 µM of STF-083010 or 3 µM of GSK2656157 for 4 h at 37°C in a 5% CO 2 incubator. T84 and Caco-2 cells were further treated with 3 µM of KIRA6 and 3 µM of GSK2656157 during C. jejuni infection or thapsigargin treatment. GAPDH was used as an internal control.

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: Western blotting showing protein levels of both spliced & unspliced forms of XBP1 [XBP1(u) & XBP1(s)] and CHOP in T84 cells infected with C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI of 200:1) or treated with 2 µM of thapsigargin (Tg) for 24 h at 37°C in a 5% CO 2 incubator. Prior to C. jejuni infection or thapsigargin treatment, T84 and Caco-2 cells were pre-treated with 3 µM of KIRA6, 100 µM of STF-083010 or 3 µM of GSK2656157 for 4 h at 37°C in a 5% CO 2 incubator. T84 and Caco-2 cells were further treated with 3 µM of KIRA6 and 3 µM of GSK2656157 during C. jejuni infection or thapsigargin treatment. GAPDH was used as an internal control.

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Western Blot, Infection, Control

(A) T84 or (B) Caco-2 cells grown in a 96-well plate were treated with 3 µM of KIRA6, 3 µM of GSK2656157 or 30 µM of ML130 for 24 h or treated with 2 µM of thapsigargin for 6 h, 100 µM of STF-083010 for 4 h or 10 µM of BAPTA-AM for 1 h at 37°C in a 5% CO 2 incubator. Medium from each well was then analysed using a LDH assay. Cytotoxicity (%) was calculated by using the following equation: ( C. jejuni infected LDH activity – spontaneous LDH activity) / (maximum LDH activity – spontaneous LDH activity) x 100. Three biological and three technical replicates were performed for each experiment. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01).

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: (A) T84 or (B) Caco-2 cells grown in a 96-well plate were treated with 3 µM of KIRA6, 3 µM of GSK2656157 or 30 µM of ML130 for 24 h or treated with 2 µM of thapsigargin for 6 h, 100 µM of STF-083010 for 4 h or 10 µM of BAPTA-AM for 1 h at 37°C in a 5% CO 2 incubator. Medium from each well was then analysed using a LDH assay. Cytotoxicity (%) was calculated by using the following equation: ( C. jejuni infected LDH activity – spontaneous LDH activity) / (maximum LDH activity – spontaneous LDH activity) x 100. Three biological and three technical replicates were performed for each experiment. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01).

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Lactate Dehydrogenase Assay, Infection, Activity Assay

(A) T84 cells in a 24-well plate were infected with C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI 200:1) or treated with 2 µM of thapsigargin for 6 or 24 h at 37°C in a 5% CO 2 incubator. T84 cells were pre-treated with 3 µM of KIRA6, 100 µM of STF-083010 or 3 µM of GSK2656157 for 4 h at 37°C in a 5% CO 2 incubator. T84 cells were then infected with C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI 200:1) or treated with 2 µM of thapsigargin for 24 h at 37°C in a 5% CO 2 incubator. T84 cells were further treated with 3 µM of KIRA6 and 3 µM of GSK2656157 during C. jejuni infection. T84 cells in a 24-well plate were infected with C. jejuni (B) 11168H, (C) 81-176 or (D) 488 wild-type strains or treated with (E) thapsigargin for 6 or 24 h at 37°C in a 5% CO 2 incubator. Medium from each well was subjected to human IL-8 ELISA to measure the concentrations of IL-8. One sample t -test was performed. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01; *** = p < 0.001; **** = p < 0.0001).

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: (A) T84 cells in a 24-well plate were infected with C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI 200:1) or treated with 2 µM of thapsigargin for 6 or 24 h at 37°C in a 5% CO 2 incubator. T84 cells were pre-treated with 3 µM of KIRA6, 100 µM of STF-083010 or 3 µM of GSK2656157 for 4 h at 37°C in a 5% CO 2 incubator. T84 cells were then infected with C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI 200:1) or treated with 2 µM of thapsigargin for 24 h at 37°C in a 5% CO 2 incubator. T84 cells were further treated with 3 µM of KIRA6 and 3 µM of GSK2656157 during C. jejuni infection. T84 cells in a 24-well plate were infected with C. jejuni (B) 11168H, (C) 81-176 or (D) 488 wild-type strains or treated with (E) thapsigargin for 6 or 24 h at 37°C in a 5% CO 2 incubator. Medium from each well was subjected to human IL-8 ELISA to measure the concentrations of IL-8. One sample t -test was performed. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01; *** = p < 0.001; **** = p < 0.0001).

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Infection, Enzyme-linked Immunosorbent Assay

qRT-PCR showing expression of human (A, C) CHOP and (B, D) spliced XBP1 [ XBP1(s) ] in T84 or Caco-2 cells infected with either C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI of 200:1) or treated with 2 µM of thapsigargin for 24 h at 37°C in a 5% CO 2 atmosphere. Prior to C. jejuni infection or thapsigargin treatment, T84 and Caco-2 cells were treated with 30 µM of ML130 for 4 h at 37°C in a 5% CO 2 incubator. T84 and Caco-2 cells were further treated with 30 µM of ML130 during C. jejuni infection or thapsigargin treatment. GAPDH was used as an internal control. (E) T84 cells were pre-treated with 30 µM of ML130 for 4 h at 37°C in a 5% CO 2 incubator. T84 cells were then infected with C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI of 200:1) or treated with 2 µM thapsigargin for 24 h at 37°C in a 5% CO 2 incubator. T84 cells were further treated with 30 µM of ML130 for 4 h during C. jejuni infection. Medium from each well was subjected to human IL-8 ELISA to measure the concentrations of IL-8. One sample t -test was performed. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01; **** = p < 0.0001).

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: qRT-PCR showing expression of human (A, C) CHOP and (B, D) spliced XBP1 [ XBP1(s) ] in T84 or Caco-2 cells infected with either C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI of 200:1) or treated with 2 µM of thapsigargin for 24 h at 37°C in a 5% CO 2 atmosphere. Prior to C. jejuni infection or thapsigargin treatment, T84 and Caco-2 cells were treated with 30 µM of ML130 for 4 h at 37°C in a 5% CO 2 incubator. T84 and Caco-2 cells were further treated with 30 µM of ML130 during C. jejuni infection or thapsigargin treatment. GAPDH was used as an internal control. (E) T84 cells were pre-treated with 30 µM of ML130 for 4 h at 37°C in a 5% CO 2 incubator. T84 cells were then infected with C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI of 200:1) or treated with 2 µM thapsigargin for 24 h at 37°C in a 5% CO 2 incubator. T84 cells were further treated with 30 µM of ML130 for 4 h during C. jejuni infection. Medium from each well was subjected to human IL-8 ELISA to measure the concentrations of IL-8. One sample t -test was performed. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01; **** = p < 0.0001).

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Quantitative RT-PCR, Expressing, Infection, Control, Enzyme-linked Immunosorbent Assay

T84 cells grown in a 96-well plate were pre-treated with 5, 10, 20 or 30 µM of BAPTA-AM for 1 h at 37°C in a 5% CO 2 incubator. T84 cells were then infected with C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI of 200:1) or treated with 2 µM thapsigargin for 24 h at 37°C in a 5% CO 2 incubator. After infection or treatment, 5 μM of cell-permeable dye Fura-2, AM was used to detect Ca 2+ . The two sets of fluorescence with 340 nm excitation and 510 nm emission (λ 1 ) and 380 nm excitation and 510 nm emission (λ 2 ) were recorded and intracellular Ca 2+ concentration was calculated . qRT-PCR showing expression of human (B) CHOP and (C) spliced XBP1 [ XBP1(s) ] in T84 cells infected with either C. jejuni 11168H wild-type strain (MOI of 200:1) or treated with 2 µM of thapsigargin for 24 h at 37°C in a 5% CO 2 atmosphere. Prior to C. jejuni infection or thapsigargin treatment, T84 cells were pre-treated with 10 µM of BAPTA-AM for 1 h at 37°C in a 5% CO 2 incubator. GAPDH was used as an internal control. One sample t -test was performed. Asterisks denote a statistically significant difference (** = p < 0.01; *** = p < 0.001; **** = p < 0.0001).

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: T84 cells grown in a 96-well plate were pre-treated with 5, 10, 20 or 30 µM of BAPTA-AM for 1 h at 37°C in a 5% CO 2 incubator. T84 cells were then infected with C. jejuni 11168H, 81-176 or 488 wild-type strains (MOI of 200:1) or treated with 2 µM thapsigargin for 24 h at 37°C in a 5% CO 2 incubator. After infection or treatment, 5 μM of cell-permeable dye Fura-2, AM was used to detect Ca 2+ . The two sets of fluorescence with 340 nm excitation and 510 nm emission (λ 1 ) and 380 nm excitation and 510 nm emission (λ 2 ) were recorded and intracellular Ca 2+ concentration was calculated . qRT-PCR showing expression of human (B) CHOP and (C) spliced XBP1 [ XBP1(s) ] in T84 cells infected with either C. jejuni 11168H wild-type strain (MOI of 200:1) or treated with 2 µM of thapsigargin for 24 h at 37°C in a 5% CO 2 atmosphere. Prior to C. jejuni infection or thapsigargin treatment, T84 cells were pre-treated with 10 µM of BAPTA-AM for 1 h at 37°C in a 5% CO 2 incubator. GAPDH was used as an internal control. One sample t -test was performed. Asterisks denote a statistically significant difference (** = p < 0.01; *** = p < 0.001; **** = p < 0.0001).

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Infection, Fluorescence, Concentration Assay, Quantitative RT-PCR, Expressing, Control

qRT-PCR showing expression of human (A) CHOP and (B) spliced XBP1 [ XBP1(s) ] in T84 cells infected with either C. jejuni 11168H wild-type strain, mutants or heat-killed 11168H wild-type strain for 24 h at 37°C in a 5% CO 2 atmosphere (MOI of 200:1). (C) RT-PCR showing expression of unspliced and spliced XBP1 [ XBP1(u) and XBP1(s) ] in T84 and Caco-2 cells infected with either C. jejuni 11168H wild-type strain or cdtABC operon mutant for 24 h at 37°C in a 5% CO 2 atmosphere. GAPDH was used as an internal control. One sample t -test was performed. Asterisks denote a statistically significant difference (** = p < 0.01; *** = p < 0.001; **** = p < 0.0001).

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: qRT-PCR showing expression of human (A) CHOP and (B) spliced XBP1 [ XBP1(s) ] in T84 cells infected with either C. jejuni 11168H wild-type strain, mutants or heat-killed 11168H wild-type strain for 24 h at 37°C in a 5% CO 2 atmosphere (MOI of 200:1). (C) RT-PCR showing expression of unspliced and spliced XBP1 [ XBP1(u) and XBP1(s) ] in T84 and Caco-2 cells infected with either C. jejuni 11168H wild-type strain or cdtABC operon mutant for 24 h at 37°C in a 5% CO 2 atmosphere. GAPDH was used as an internal control. One sample t -test was performed. Asterisks denote a statistically significant difference (** = p < 0.01; *** = p < 0.001; **** = p < 0.0001).

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Quantitative RT-PCR, Expressing, Infection, Reverse Transcription Polymerase Chain Reaction, Mutagenesis, Control

Interactions with and invasion of T84 intestinal epithelial cells by C. jejuni 11168H wild-type strain and different mutants . T84 cells were infected with C. jejuni 11168H wild-type strain, 11168H mutants or heat-killed 11168H wild-type strain for 3 h at 37°C in a 5% CO 2 incubator (MOI of 200:1). (A) For interaction assays, T84 cells were then washed with PBS and lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. (B) For invasion assays, T84 cells were then incubated with gentamicin (150 µg/ml) for 2 h to kill extracellular bacteria and then lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. Three biological and three technical replicates were performed for each experiment. # denotes no growth was observed. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01; *** = p < 0.001).

Journal: bioRxiv

Article Title: Unfolded protein response is a critical mediator in Campylobacter jejuni pathogenesis and host defence

doi: 10.1101/2025.01.31.635839

Figure Lengend Snippet: Interactions with and invasion of T84 intestinal epithelial cells by C. jejuni 11168H wild-type strain and different mutants . T84 cells were infected with C. jejuni 11168H wild-type strain, 11168H mutants or heat-killed 11168H wild-type strain for 3 h at 37°C in a 5% CO 2 incubator (MOI of 200:1). (A) For interaction assays, T84 cells were then washed with PBS and lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. (B) For invasion assays, T84 cells were then incubated with gentamicin (150 µg/ml) for 2 h to kill extracellular bacteria and then lysed with 0.1% (v/v) Triton X-100 and CFU/ml were recorded after incubation. Three biological and three technical replicates were performed for each experiment. # denotes no growth was observed. Asterisks denote a statistically significant difference (* = p < 0.05; ** = p < 0.01; *** = p < 0.001).

Article Snippet: Human carcinoma cell line T84 cells (ECACC 88021101) and Caco-2 cells (ECACC 86010202) were obtained from European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Infection, Incubation, Bacteria