Journal: bioRxiv

Article Title: RodA Promotes Intestinal Colonization by Group B Streptococcus

doi: 10.1101/2025.10.27.684869

Figure Lengend Snippet: C. Adhesion assays were performed with Caco-2 and T84 intestinal cell lines respectively at an MOI of 1 to 10. B, D. Invasion assays were performed with Caco-2 and T84 intestinal cell lines respectively at an MOI of 1 to 10. The percentage of adhered and invasive bacteria were calculated based on the initial inoculum and expressed relative to the value for the WT strain. All data are from independent experiments (n=9, each with 3 technical replicates). The error bars indicate the 95% confidence intervals of the means of the wells. Data were analyzed using the Mann-Whitney U test (****, p < 0.0001) .

Article Snippet: The T84 colonic epithelial cell line (ATCC CCL-248) were cultivated in Dulbecco’s Modified Eagle’s Medium and Ham F-12 medium (DMEM: F-12, ATCC 30-2006) supplemented with 10% FBS and 1% Pen/Strep ( ).

Techniques: Bacteria, MANN-WHITNEY

Journal: Food Hydrocolloids for Health

Article Title: Pectins mitigate Candidalysin-induced gut barrier disruption and inflammation in vitro in a degree of methyl-esterification-dependent manner

doi: 10.1016/j.fhfh.2025.100232

Figure Lengend Snippet: Fig. 1. Treatment schedule and experimental design. T84 cells were seeded in ECIS plates until form a differentiated monolayer. Then, cells were pre-treated with 2 mg/mL of DM18 or DM88 pectin for 24 h (A). The next day, cells were exposed or to 10 µM CaLysin for 24 h (B). Untreated cells (complete medium with the vehicle) were used as negative control.

Article Snippet: To study the protective effects of lemon pectins against CaLysin barrier disruption and the expression of TJs-, inflammation- and wound healing-related genes, the human colorectal carcinoma epithelial cell line T84 (ATCC® CCL-248TM, Manassas, USA) was used in all the experiments.

Techniques: Negative Control

Journal: Food Hydrocolloids for Health

Article Title: Pectins mitigate Candidalysin-induced gut barrier disruption and inflammation in vitro in a degree of methyl-esterification-dependent manner

doi: 10.1016/j.fhfh.2025.100232

Figure Lengend Snippet: Fig. 2. The protective effect of DM18 and DM88 pectins on T84 epithelial intestinal barrier disruption induced by Candidalysin (CaLysin). Confluent monolayers of T84 cells were pre-incubated with 2 mg/mL of DM18 or DM88 for 24 h. Afterwards, the fungal toxin CaLysin (10 μM) was added, and the cells were incubated for 24 h. (A) The representative figure of barrier function measurement with ECIS system. (B) Calculated AUC. The % AUC of the T84 cells pre-incubated with only pectins were compared with the untreated controls. Statistical differences between the positive control and the different conditions were determined. Data from four independent experiments are presented as mean ± SEM. ** (p < 0.01) vs. Control or ## (p < 0.01) vs. CaLysin.

Article Snippet: To study the protective effects of lemon pectins against CaLysin barrier disruption and the expression of TJs-, inflammation- and wound healing-related genes, the human colorectal carcinoma epithelial cell line T84 (ATCC® CCL-248TM, Manassas, USA) was used in all the experiments.

Techniques: Disruption, Incubation, Positive Control, Control

Journal: Food Hydrocolloids for Health

Article Title: Pectins mitigate Candidalysin-induced gut barrier disruption and inflammation in vitro in a degree of methyl-esterification-dependent manner

doi: 10.1016/j.fhfh.2025.100232

Figure Lengend Snippet: Fig. 3. Pectin-mediated cytoprotection after CaLysin challenge. Determination of LDH (A) and ALDH (B) release in T84 cells expressed as Percent cytotoxicity. Data from four independent experiments are presented as mean ± SEM. * (p < 0.05), **** (p < 0.0001) vs. Control or # (p < 0.05), #### (p < 0.0001) vs. CaLysin.

Article Snippet: To study the protective effects of lemon pectins against CaLysin barrier disruption and the expression of TJs-, inflammation- and wound healing-related genes, the human colorectal carcinoma epithelial cell line T84 (ATCC® CCL-248TM, Manassas, USA) was used in all the experiments.

Techniques: Control

Journal: Molecular Nutrition & Food Research

Article Title: Protective Effects of Human Milk Oligosaccharides (hMOs) on Analgesic, Nonsteroidal Anti‐inflammatory Drugs (NSAIDs)‐Induced Disruption of Gut Barrier Integrity in Cocultures Mimicking Neuro‐Gut Epithelial Cell Crosstalk

doi: 10.1002/mnfr.70146

Figure Lengend Snippet: The impact of SH‐SY5Y nerve cells on indomethacin‐induced disruption of intestinal epithelial barrier for Peer Review after 2 weeks of growth, 280 µM indomethacin was applied to disrupt the confluent monolayers for 24 h. T84 cells without indomethacin were used as negative controls and indomethacin‐induced T84 cells were used as positive controls. The change in TEER after the addition of indomethacin in monolayers compared to the untreated controls was illustrated in (A). The AUC was calculated as shown in (B). The AUC of untreated control was set to 100%. The % AUC of the indomethacin‐induced disruption in the cocultures was compared with positive control. Statistical significance was determined between the untreated control and positive control, and between the positive control and different conditions. Data are displayed as mean ± SEM based on a minimum of five independent experiments. #### p < 0.0001.

Article Snippet: T84 colorectal carcinoma cell line (ATCC CCL‐248, Manassas, USA) between Passages 12–22 and SH‐SY5Y human neuroblastoma cell line (ATCC CRL‐2266, Manassas, USA) between Passages 52–62 were used in all experiments.

Techniques: Disruption, Control, Positive Control

Journal: Molecular Nutrition & Food Research

Article Title: Protective Effects of Human Milk Oligosaccharides (hMOs) on Analgesic, Nonsteroidal Anti‐inflammatory Drugs (NSAIDs)‐Induced Disruption of Gut Barrier Integrity in Cocultures Mimicking Neuro‐Gut Epithelial Cell Crosstalk

doi: 10.1002/mnfr.70146

Figure Lengend Snippet: The impact of SH‐SY5Y cells on the expression of genes involved in modulating the gut barrier in T84/SH‐SY5Y cocultures. The cocultures were incubated with 280 µM indomethacin for 24 h. The T84 cells without any conditions were the untreated control, and the T84 cells incubated with indomethacin were the positive control. qPCR was used to identify the expression of the following genes: CLDN1 (A), CLDN2 (B), CLDN3 (C), CLDN4 (D), OCLN (E), ZO‐1 (F), VIP (G), and NPY (H). Statistical significance was determined between the untreated control and positive control, and between the positive control and different conditions. Data are displayed as mean ± SEM based on a minimum of five independent experiments. * p < 0.05, ## or ** p < 0.01, #### or **** p < 0.0001.

Article Snippet: T84 colorectal carcinoma cell line (ATCC CCL‐248, Manassas, USA) between Passages 12–22 and SH‐SY5Y human neuroblastoma cell line (ATCC CRL‐2266, Manassas, USA) between Passages 52–62 were used in all experiments.

Techniques: Expressing, Incubation, Control, Positive Control

Journal: Molecular Nutrition & Food Research

Article Title: Protective Effects of Human Milk Oligosaccharides (hMOs) on Analgesic, Nonsteroidal Anti‐inflammatory Drugs (NSAIDs)‐Induced Disruption of Gut Barrier Integrity in Cocultures Mimicking Neuro‐Gut Epithelial Cell Crosstalk

doi: 10.1002/mnfr.70146

Figure Lengend Snippet: Influence of SH‐SY5Y nerve cells on claudin‐1 and VIP protein expression in T84 cells in cocultures of gut epithelial and nerve cells at ratios of 29:1 and 14:1 after 2 weeks of culture. The cells were treated with 280 µM indomethacin for 24 h. Indomethacin‐induced disrupted T84 monolayers were used as the positive control for comparing claudin‐1 expression in both T84 cells and cocultures. Similarly, A23187‐induced T84:SH‐SY5Y = 29:1 coculture served as the positive control for comparing VIP expression in the cocultures. Immunofluorescence staining identified claudin‐1 ((A) green; cell nuclei, blue) and VIP ((B) red; cell nuclei, blue); scale bar = 25 µm. The immunofluorescence intensity levels of claudin‐1 and VIP were quantified by ImageJ. Statistical comparisons between the untreated control and the positive control; and between the positive control and different conditions were calculated. Immunofluorescence intensity data are presented as mean ± SEM ( n = 6). ** p < 0.01.

Article Snippet: T84 colorectal carcinoma cell line (ATCC CCL‐248, Manassas, USA) between Passages 12–22 and SH‐SY5Y human neuroblastoma cell line (ATCC CRL‐2266, Manassas, USA) between Passages 52–62 were used in all experiments.

Techniques: Expressing, Positive Control, Immunofluorescence, Staining, Control

Journal: Molecular Nutrition & Food Research

Article Title: Protective Effects of Human Milk Oligosaccharides (hMOs) on Analgesic, Nonsteroidal Anti‐inflammatory Drugs (NSAIDs)‐Induced Disruption of Gut Barrier Integrity in Cocultures Mimicking Neuro‐Gut Epithelial Cell Crosstalk

doi: 10.1002/mnfr.70146

Figure Lengend Snippet: The effect of 2′‐FL and 3‐FL on indomethacin disrupted the intestinal epithelial barrier in the presence and absence of nerve cells. T84 intestinal epithelial cells were cocultured with SH‐SY5Y nerve cells in ratios of 29:1 and 14:1. After 2 weeks of growth, confluent monolayers were preincubated with 2 mg/mL 2′‐FL or 3‐FL for 24 h. Then 280 µM indomethacin was applied to disrupt the barrier for 24 h. Monolayers without any exposure to barrier disruptors were used as negative controls, and indomethacin‐induced disrupted monolayers were used as positive controls. The relative TEER change after the addition of indomethacin in T84, T84:SH‐SY5Y = 29:1, and T84:SH‐SY5Y = 14:1 monolayers are illustrated in (A), (C), and (E), respectively. The AUC was calculated as shown in (B), (D), and (F), respectively. The AUC of untreated control was set to 100%. The % AUC of hMOs preincubated and indomethacin‐stimulated monolayers was compared with that of the untreated control. Statistical significance was determined between the untreated control and positive control, and between the positive control and different conditions. Data are displayed as mean ± SEM based on a minimum of five independent experiments. * p < 0.05, #### p < 0.0001.

Article Snippet: T84 colorectal carcinoma cell line (ATCC CCL‐248, Manassas, USA) between Passages 12–22 and SH‐SY5Y human neuroblastoma cell line (ATCC CRL‐2266, Manassas, USA) between Passages 52–62 were used in all experiments.

Techniques: Control, Positive Control

Journal: Molecular Nutrition & Food Research

Article Title: Protective Effects of Human Milk Oligosaccharides (hMOs) on Analgesic, Nonsteroidal Anti‐inflammatory Drugs (NSAIDs)‐Induced Disruption of Gut Barrier Integrity in Cocultures Mimicking Neuro‐Gut Epithelial Cell Crosstalk

doi: 10.1002/mnfr.70146

Figure Lengend Snippet: The impact of 2′‐FL and 3‐FL on the TJs genes of T84 cells. 2 mg/mL 2′‐FL or 3‐FL was first added to the T84 cells and incubated for 24 h, then the T84 cells were challenged with 280 µM indomethacin 24 h. The T84 cells without any conditions were the untreated control, and the T84 cells incubated with indomethacin were the positive control. qPCR was used to identify the expression of the following genes: CLDN1 (A), CLDN2 (B), CLDN3 (C), CLDN4 (D), OCLN (E), and ZO‐1 (F). Statistical significance was determined between the untreated control and positive control, and between the positive control and different conditions. Data are displayed as mean ± SEM based on a minimum of five independent experiments. * p < 0.05, ** p < 0.01, #### p < 0.0001.

Article Snippet: T84 colorectal carcinoma cell line (ATCC CCL‐248, Manassas, USA) between Passages 12–22 and SH‐SY5Y human neuroblastoma cell line (ATCC CRL‐2266, Manassas, USA) between Passages 52–62 were used in all experiments.

Techniques: Incubation, Control, Positive Control, Expressing

Journal: Molecular Nutrition & Food Research

Article Title: Protective Effects of Human Milk Oligosaccharides (hMOs) on Analgesic, Nonsteroidal Anti‐inflammatory Drugs (NSAIDs)‐Induced Disruption of Gut Barrier Integrity in Cocultures Mimicking Neuro‐Gut Epithelial Cell Crosstalk

doi: 10.1002/mnfr.70146

Figure Lengend Snippet: The impact of 2′‐FL and 3‐FL on the expression of genes involved in regulating the gut barrier in a coculture of T84 and SH‐SY5Y cells at a 14:1 ratio. The hMOs 2′‐FL or 3‐FL were first added to the coculture and incubated for 24 h, then the coculture was challenged with 280 µM indomethacin for 24 h. The coculture without any conditions was the untreated control, and the coculture incubated with indomethacin was the positive control. qPCR was used to identify the expression of the following genes: CLDN1 (A), CLDN2 (B), CLDN3 (C), CLDN4 (D), OCLN (E), ZO‐1 (F), VIP (G), and NPY (H). Statistical significance was determined between the untreated control and positive control, and between the positive control and different conditions. Data are displayed as mean ± SEM based on a minimum of five independent experiments. # or * p < 0.05, ** p < 0.01, #### or **** p < 0.0001.

Article Snippet: T84 colorectal carcinoma cell line (ATCC CCL‐248, Manassas, USA) between Passages 12–22 and SH‐SY5Y human neuroblastoma cell line (ATCC CRL‐2266, Manassas, USA) between Passages 52–62 were used in all experiments.

Techniques: Expressing, Incubation, Control, Positive Control

Journal: Molecular Nutrition & Food Research

Article Title: Protective Effects of Human Milk Oligosaccharides (hMOs) on Analgesic, Nonsteroidal Anti‐inflammatory Drugs (NSAIDs)‐Induced Disruption of Gut Barrier Integrity in Cocultures Mimicking Neuro‐Gut Epithelial Cell Crosstalk

doi: 10.1002/mnfr.70146

Figure Lengend Snippet: Effects of 2′‐FL and 3‐FL on claudin‐1 and VIP protein expression in indomethacin‐disrupted T84 cells (A, B), as well as in cocultures of nerve cells and epithelial cells at ratios of 29:1 (C, D) and 14:1 (E, F) after 2 weeks of culture. The cells were treated with 2 mg/mL of 2′‐FL and 3‐FL for 24 h, followed by a disruption with 280 µM indomethacin for another 24 h. Indomethacin‐treated T84 cells and indomethacin‐exposed cocultures were used as positive controls. Immunofluorescence staining identified cell nuclei (blue), claudin‐1 (green), VIP (red), and F‐actin (purple). Arrows indicate examples of F‐actin structures in cells under different conditions. Scale bar = 25 µm. The immunofluorescence intensity levels of claudin‐1 and VIP were quantified by ImageJ. Statistical comparisons between the untreated control and the positive control; and between the positive control and different conditions were calculated. Immunofluorescence intensity data are presented as mean ± SEM ( n = 6). * p < 0.05, #### or **** p < 0.0001.

Article Snippet: T84 colorectal carcinoma cell line (ATCC CCL‐248, Manassas, USA) between Passages 12–22 and SH‐SY5Y human neuroblastoma cell line (ATCC CRL‐2266, Manassas, USA) between Passages 52–62 were used in all experiments.

Techniques: Expressing, Disruption, Immunofluorescence, Staining, Control, Positive Control