Journal: Biomolecules

Article Title: Vitamin D and Retinoic Acid Require Protein Kinase C Activity and Reactive Oxygen Species as Opposing Signals Regulating PEIG-1 / GPRC5A Expression in Caco-2 and T84 Colon Carcinoma Cells.

doi: 10.3390/biom15050711

Figure Lengend Snippet: Figure 1. VD and RA increase GPRC5A mRNA expression. (A) GPRC5A mRNA expression levels in Caco-2 and T84 cell lines, following RA (0.1–10 µM) treatment for 4 h. (B) GPRC5A mRNA expression levels in Caco-2 and T84 cell lines following VD (1–40 µM) treatment for 24 h (C) GPRC5A mRNA expression levels after combined RA and VD treatment for 4 or (D) 24 h. Data are expressed as mean ± SEM (n) for three independent experiments (n = 3). Each dot represents the mean value of each independent experiment. Statistical analyses were performed using ANOVA followed by Tukey’s post hoc tests. (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p <0.0001; ## p < 0.01, ### p < 0.001). All figures regarding expression levels represent fold changes compared to control values.

Article Snippet: Human colon adenocarcinoma cell lines T84 (CCL-248) and Caco-2 (HTB-37) were purchased from the American Type Culture Collection (ATCC).

Techniques: Expressing, Control

Journal: Biomolecules

Article Title: Vitamin D and Retinoic Acid Require Protein Kinase C Activity and Reactive Oxygen Species as Opposing Signals Regulating PEIG-1 / GPRC5A Expression in Caco-2 and T84 Colon Carcinoma Cells.

doi: 10.3390/biom15050711

Figure Lengend Snippet: Figure 2. Modulation of GPRC5A gene expression. (A) Changes in GPRC5A mRNA expression levels in Caco-2 (left panel) and T84 (right panel) cell lines, in the presence or absence of VD (10 µM) and Gö6983 (10 µM), for 24 h. (B) Changes in GPRC5A mRNA expression levels in Caco-2 and T84 cell lines, in the presence or absence of RA (10 µM) and Gö6983 (10 µM), for 4 h (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; # p < 0.05; ### p < 0.001; #### p < 0.0001).

Article Snippet: Human colon adenocarcinoma cell lines T84 (CCL-248) and Caco-2 (HTB-37) were purchased from the American Type Culture Collection (ATCC).

Techniques: Gene Expression, Expressing

Journal: Biomolecules

Article Title: Vitamin D and Retinoic Acid Require Protein Kinase C Activity and Reactive Oxygen Species as Opposing Signals Regulating PEIG-1 / GPRC5A Expression in Caco-2 and T84 Colon Carcinoma Cells.

doi: 10.3390/biom15050711

Figure Lengend Snippet: Figure 3. VD and RA induce changes in ROS levels. Changes in cytoplasmic (DCFH) and mito- chondrial ROS (MitoSOX) fluorescence levels in Caco-2 cells (left panels) and T84 cells (right panels) treated with vitamin D (VD) or retinoic acid (RA). (A) cROS levels in the presence of VD (10 µM) for 24 h. (B) mtROS levels in the presence of VD (10 µM) for 24 h. (C) cROS levels in the presence of RA (10 µM) for 4 h. (D) mtROS levels in the presence of RA (10 µM) for 4 h (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

Article Snippet: Human colon adenocarcinoma cell lines T84 (CCL-248) and Caco-2 (HTB-37) were purchased from the American Type Culture Collection (ATCC).

Techniques: Fluorescence

Journal: Biomolecules

Article Title: Vitamin D and Retinoic Acid Require Protein Kinase C Activity and Reactive Oxygen Species as Opposing Signals Regulating PEIG-1 / GPRC5A Expression in Caco-2 and T84 Colon Carcinoma Cells.

doi: 10.3390/biom15050711

Figure Lengend Snippet: Figure 4. Modulation of GPRC5A gene expression in the presence of ROS scavengers. (A) GPRC5A mRNA expression levels in Caco-2 and T84 cell lines, in the presence/absence of VD (10 µM) and NAC (10 mM). (B) GPRC5A mRNA expression levels in Caco-2 and T84 cell lines, in the presence/absence of RA (10 µM) and NAC (10 mM). (C) GPRC5A mRNA expression levels in Caco-2 and T84 cell lines, in the presence/absence of VD (10 µM) and MitoTEMPO (10 µM). (D) GPRC5A mRNA expression levels in Caco-2 and T84 cell lines, in the presence/absence of RA (10 µM) and MitoTEMPO (10 µM). The strongest response was observed with MitoTEMPO and RA (* p < 0.05; ** p < 0.01; *** p < 0.001; # p < 0.05; ## p < 0.01; #### p < 0.0001).

Article Snippet: Human colon adenocarcinoma cell lines T84 (CCL-248) and Caco-2 (HTB-37) were purchased from the American Type Culture Collection (ATCC).

Techniques: Gene Expression, Expressing

Journal: Experimental and molecular pathology

Article Title: Cross-feeding between beneficial and pathogenic bacteria to utilize eukaryotic host cell-derived sialic acids and bacteriophages shape the pathogen-host interface milieu.

doi: 10.1016/j.yexmp.2025.104967

Figure Lengend Snippet: Fig. 1. Cell viability assay. The impact of P.a PAO1, KPP22 phage, and B. bifidum on the cell viability of T84 cells (panel A), THP-1 cells (panel B), and Huh7 cells (panel C) is shown. The seeded triple co-cultured cells were incubated in DMEM culture medium (white bars) or DMEM culture medium with 100 μM of DANA (black bars). The cells were then exposed for 6 h to the following treatments: 1) P.a PAO1 (1 × 108CFU/ml) alone; 2) P.a PAO1 plus its KPP22 phage; and 3) P.a PAO1 plus B. bifidum (1 × 108CFU/ml). In the control group, cells were incubated with only medium. The KPP22 phage was used as a specific natural phage lysing P.a PAO1. DANA was used as a specific endogenous sialidase inhibitor. The mean values of the control wells containing only cells without any bacteria were taken as 100 %. The percentage of viable active cells treated with bacteria, phage, or DANA was then calculated. Each bar represents the mean of three independent replicates (n = 3) with its corresponding standard error of the mean (SEM). Statistical comparisons were conducted using a one-way ANOVA with Tukey’s post-test (A, B, and C). Data are expressed as percentage (%) cell survival compared to the control. The bars with different lowercase letters (a-g) indicate statistically significant differences between any two groups (p < 0.05), indicating that they are significantly different from each other. Bars with the same lowercase letters represent no significant differences between any two groups (p < 0.05). P.a PAO1, Pseudomonas aeruginosa PAO1; B.bifidum, Bifidobacterium bifidum; KPP22, KPP22 phage.

Article Snippet: The human colon carcinoma cell line (T84) and the human monocyte cell line THP-1 (cell stocks frozen in liquid nitrogen) were from the American Type Culture Collection (ATCC, LGC, Wesel, Germany).

Techniques: Viability Assay, Cell Culture, Incubation, Control, Bacteria

Journal: Experimental and molecular pathology

Article Title: Cross-feeding between beneficial and pathogenic bacteria to utilize eukaryotic host cell-derived sialic acids and bacteriophages shape the pathogen-host interface milieu.

doi: 10.1016/j.yexmp.2025.104967

Figure Lengend Snippet: Fig. 2. Transepithelial electrical resistance (TEER) of T84 cell monolayers. The monolayers of T84 cells grown on transwell inserts were exposed to P.a PAO1 (1 × 108 CFU/ml) and B. bifidum (1 × 108 CFU/ml) for 6 h. The error bars indicate the standard deviation of at least three independent experiments. Data are expressed as means ±SEM. Bars with different lowercase letters represent statistically significant differences between any two groups (p < 0.05), indi cating that they are significantly different from each other. The bars with common (same) lowercase letters represent no significant differences between any two groups (p < 0.05).

Article Snippet: The human colon carcinoma cell line (T84) and the human monocyte cell line THP-1 (cell stocks frozen in liquid nitrogen) were from the American Type Culture Collection (ATCC, LGC, Wesel, Germany).

Techniques: Standard Deviation

Journal: Experimental and molecular pathology

Article Title: Cross-feeding between beneficial and pathogenic bacteria to utilize eukaryotic host cell-derived sialic acids and bacteriophages shape the pathogen-host interface milieu.

doi: 10.1016/j.yexmp.2025.104967

Figure Lengend Snippet: Fig. 3. Release of free sialic acid from the surface of co-cultured cells and sialidase activity in the cell culture supernatant. T84, THP-1, and Huh7 cells were co-cultured in DMEM medium or DMEM + DANA and incubated with P.a PAO1, P.a PAO1+ KPP22 phage or P.a PAO1+ B. bifidum for 6 h. The control group was incubated with only medium for comparison. (A) The amount of free sialic acid remaining in the cell culture supernatants 6 h after inoculation of P.a PAO1 and B. bifidum or uninoculated media control. (B) Sialidase activity was measured in 6-h co-cultures using supernatants from panel A. After 6 h of in cubation, the activity of sialidase in the supernatants was analyzed by spec trophotometry and expressed as absorbance (Ab) at 595 nm. Both panels show the mean values of three independent experiments, with standard errors of the mean (± SEM). The bars with different lowercase letters (a-g) represent sta tistically significant differences between any two groups (p < 0.05), indicating that they differ significantly from each other. The bars with common (same) lowercase letters represent no significant differences between any two groups (p < 0.05). The bars represent the mean values of three independent experi ments carried out in duplicate.

Article Snippet: The human colon carcinoma cell line (T84) and the human monocyte cell line THP-1 (cell stocks frozen in liquid nitrogen) were from the American Type Culture Collection (ATCC, LGC, Wesel, Germany).

Techniques: Cell Culture, Activity Assay, Incubation, Control, Comparison

Journal: Experimental and molecular pathology

Article Title: Cross-feeding between beneficial and pathogenic bacteria to utilize eukaryotic host cell-derived sialic acids and bacteriophages shape the pathogen-host interface milieu.

doi: 10.1016/j.yexmp.2025.104967

Figure Lengend Snippet: Fig. 4. Influence of DANA, the KPP22 phage, and B. bifidum on the host- pathogen interaction of P.a PAO1 with T84 monolayer cells. The effects of KPP22 phage (1 × 108 PFU/ml), B. bifidum (1 × 10 8 PFU/ml), and the sialidase inhibitor DANA on P.a PAO1 (1 × 108CFU/ml) adhesion to (A) and invasion into (B) human T84 intestinal epithelial cells after 6h of infection. The error bars indicate the standard deviation of at least three independent experiments. Data are expressed as means ±SEM. The bars with different lowercase letters represent statistically significant differences between any two groups (p < 0.05), indicating that they differ significantly from each other. The bars with common (same) lowercase letters represent no significant differences between any two groups (p < 0.05).

Article Snippet: The human colon carcinoma cell line (T84) and the human monocyte cell line THP-1 (cell stocks frozen in liquid nitrogen) were from the American Type Culture Collection (ATCC, LGC, Wesel, Germany).

Techniques: Infection, Standard Deviation

Journal: Pharmaceutics

Article Title: Magnetic Nanoparticles with On-Site Azide and Alkyne Functionalized Polymer Coating in a Single Step through a Solvothermal Process

doi: 10.3390/pharmaceutics16091226

Figure Lengend Snippet: Hyperthermia and in vitro proliferation analyses: ( A ) Temperature rise obtained after application of an alternating magnetic field (385 kHz; 28 kA/m) during 25 min for MNPs coated with PEG400, PEG6000, and βCD at a concentration of 0.5 mg/mL of Fe. The data were represented as the mean of 3 measurements ± standard deviation; ( B ) In vitro proliferation assay of MNPs coated with PEG400, PEG6000, and βCD at 72 h of exposition. Graphs show the percentage of proliferation of T84 at doses ranging from 1–100 µg/mL of Fe 3 O 4 . Results were expressed as mean ± SD of triplicate cultures.

Article Snippet: The human colon cancer cell line T84 was obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: In Vitro, Concentration Assay, Standard Deviation, Proliferation Assay

Journal: Archives of toxicology

Article Title: Urolithin A attenuates arsenic-induced gut barrier dysfunction

doi: 10.1007/s00204-022-03232-2

Figure Lengend Snippet: UroA protects against iAs3+ induced cytotoxicity in colon epithelial cells. T84 cells (2 × 104) per well were grown O/N in 96 well plate. Next day, cells were treated with iAs3+ (0.1, 0.3, 1, 3, 10, 30, 100 μM) in the presence of UroA (25 μM) or vehicle (DMSO, 0.05%) for (A) 24 h, (B) 48 h and (C) 72 h. Cell viability was determined using alamarBlue assay. Percent control cell viability against iAs3+ dose were plotted (log(inhibitor) vs. normalized response (variable slope) curve fit). **p < 0.01, ***p < 0.001, Two-way Repeated Measures ANOVA with Geisser-Greenhouse’s Correction followed by Tukey’s Multiple Comparisons Test between iAs3+ and iAs3++UroA. Each point in each curve represents the mean ± SD from three independent experiments.

Article Snippet: The human colon epithelial carcinoma cell line T84 (ATCC # CCL-248 TM )) was maintained in DMEM: F-12 Medium (Cytiva # SH30261.01), supplemented with 10% fetal bovine serum, 1X penicillin-streptomycin solution (100 U/ml penicillin, and 100 μg/ml streptomycin (Sigma Aldrich) in a humidified atmosphere (at 37 °C in a 5% CO 2 incubator).

Techniques: Alamar Blue Assay, Control

Journal: Archives of toxicology

Article Title: Urolithin A attenuates arsenic-induced gut barrier dysfunction

doi: 10.1007/s00204-022-03232-2

Figure Lengend Snippet: (A) T84 cells were treated with iAs+3 (0, 1, 5, 10, 50, 100 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) for 48 h. Bar graph and images representing apoptosis in T84 cells after the indicated treatment estimated by Annexin V/PI assay. (B) Representative fluorescence image of JC1 stained T84 cells representing the effect of iAs+3 in presence of vehicle or UroA on mitochondrial permeability (red to the green shift of fluorescence). Bar graphs showing the detection of JC-1 fluorescence as J aggregates (red) vs J monomers (green). Results are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, Statistics performed using 2way ANOVA in GraphPad Prism software. Error bar, mean ± SEM (n=4–5)

Article Snippet: The human colon epithelial carcinoma cell line T84 (ATCC # CCL-248 TM )) was maintained in DMEM: F-12 Medium (Cytiva # SH30261.01), supplemented with 10% fetal bovine serum, 1X penicillin-streptomycin solution (100 U/ml penicillin, and 100 μg/ml streptomycin (Sigma Aldrich) in a humidified atmosphere (at 37 °C in a 5% CO 2 incubator).

Techniques: Fluorescence, Staining, Permeability, Software

Journal: Archives of toxicology

Article Title: Urolithin A attenuates arsenic-induced gut barrier dysfunction

doi: 10.1007/s00204-022-03232-2

Figure Lengend Snippet: (A) T84 cells were treated with iAs+3 (0, 0.05, 0.1, 1, 5, 10, 50, 100 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) for 12 h. Representative fluorescence image and bar graph are showing ROS generation as green fluorescence from DCFDA stained T84 cells. (B) LDH release after 24 h incubation of T84 cells with iAs+3 (5, 10, 20 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) (C). T-84 cells were treated with UroA (10, 25 μM). T84 cells were treated with iAs+3 (10 μM) and different concentration of UroA (0, 0.01, 0.1, 1, 5, 10, 25 μM) for 24 h. The levels of GSH and GSSG were measured and the GSH/GSSG ratio was calculated. Untreated (UT) cells were used as control (100%). Results are representative of three independent experiments. Statistics performed using 2way ANOVA in GraphPad Prism software. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Error bar, mean ± SEM (n=4–8)

Article Snippet: The human colon epithelial carcinoma cell line T84 (ATCC # CCL-248 TM )) was maintained in DMEM: F-12 Medium (Cytiva # SH30261.01), supplemented with 10% fetal bovine serum, 1X penicillin-streptomycin solution (100 U/ml penicillin, and 100 μg/ml streptomycin (Sigma Aldrich) in a humidified atmosphere (at 37 °C in a 5% CO 2 incubator).

Techniques: Fluorescence, Staining, Incubation, Concentration Assay, Control, Software

Journal: Archives of toxicology

Article Title: Urolithin A attenuates arsenic-induced gut barrier dysfunction

doi: 10.1007/s00204-022-03232-2

Figure Lengend Snippet: (A) Schematic representation of in vitro permeability study with T84 monolayers (B) and (C) Monolayer T84 cells on transmembrane were treated with iAs+3 (0, 1, 5, 10, 20 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) for 24 h. FITC-dextran was added to these cells (top of the membrane) and incubated for 2 h at 37°C and FITC-dextran levels in the bottom chamber well was measured. TEER values were also measured. (B) LDH release after 24 h incubation of T84 monolayer cells with iAs+3 (5, 10, 20 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) (C) T84 monolayer cells were incubated with iAs+3 (5, 10 μM) without or with UroA (0, 10, 25 μM) for 24 h. IL-8 levels in supernatants were measured. *p < 0.05, **p < 0.01, ***p < 0.001, Statistics performed using 2way ANOVA in GraphPad Prism software. Error bar, mean ± SEM (n=4).

Article Snippet: The human colon epithelial carcinoma cell line T84 (ATCC # CCL-248 TM )) was maintained in DMEM: F-12 Medium (Cytiva # SH30261.01), supplemented with 10% fetal bovine serum, 1X penicillin-streptomycin solution (100 U/ml penicillin, and 100 μg/ml streptomycin (Sigma Aldrich) in a humidified atmosphere (at 37 °C in a 5% CO 2 incubator).

Techniques: In Vitro, Permeability, Membrane, Incubation, Software

Journal: Archives of toxicology

Article Title: Urolithin A attenuates arsenic-induced gut barrier dysfunction

doi: 10.1007/s00204-022-03232-2

Figure Lengend Snippet: T84 cells were treated with iAs+3 (0, 5, 10 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) for 24 h. (A) Protein expression of Zona occludens 1 (ZO1), claudin 4 (Cldn4) and occludin (Ocln) in T84 cells were determined by immunoblots. (B) The fold changes in mRNA levels of ZO1, Cldn4, Ocln, and ZO1 in T84 cells were determined by RT PCR method. (C) T84 cells were grown on chambered slides and treated with iAs+3 (10 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) for 24 h. The cells were stained with rabbit anti ZO-1, rabbit anti Ocln and mouse anti-Cldn4, followed by secondary antibody tagged with anti-rabbit Alexa 488 for ZO-1, Ocln and anti-mouse Alexa-594 for cldn-4. Nucleus was stained using DAPI. The confocal images were captured. Scale bars for T84 cells indicate 20 μm respectively. The fluorescence intensity (n = 15–20 cell membrane regions) was measured. Results are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, Statistics performed using one-way ANOVA in GraphPad Prism software. Error bar, mean ± SEM (n=3–5).

Article Snippet: The human colon epithelial carcinoma cell line T84 (ATCC # CCL-248 TM )) was maintained in DMEM: F-12 Medium (Cytiva # SH30261.01), supplemented with 10% fetal bovine serum, 1X penicillin-streptomycin solution (100 U/ml penicillin, and 100 μg/ml streptomycin (Sigma Aldrich) in a humidified atmosphere (at 37 °C in a 5% CO 2 incubator).

Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Staining, Fluorescence, Membrane, Software