Journal: Frontiers in Nutrition

Article Title: Enhancement of intestinal barrier function and alleviation of mycophenolic acid toxicity by a probiotic-conditioned medium in vitro

doi: 10.3389/fnut.2026.1809197

Figure Lengend Snippet: Effect of probiotic-conditioned (PP), Mycophenolic acid (MPA), and mycophenolate mofetil (MMF) on transepithelial electrical resistance (TEER) of differentiated CaCo-2 cells. The bar graph shows the mean and standard deviation (SD) of values measured in six experiments performed with differentiated CaCo-2 cells exposed to either MMF or MPA (both at a concentration of 50 μM) or their vehicle in normal cell culture medium or PP for 6 h. Data are expressed as a percentage of the control group. Asterisks indicate statistical significance after the ANOVA (multiple comparison) analysis: ** p < 0.005; *** p < 0.001; **** p < 0.0001.

Article Snippet: Mycophenolate mofetil (MMF) and mycophenolic acid (MPA), produced by the Cayman Chemical Company (Ann Arbor, MI, United States), were obtained from Cabru S.A.S. (Arcore, Milan, Italy).

Techniques: Standard Deviation, Concentration Assay, Cell Culture, Control, Comparison

Journal: Frontiers in Nutrition

Article Title: Enhancement of intestinal barrier function and alleviation of mycophenolic acid toxicity by a probiotic-conditioned medium in vitro

doi: 10.3389/fnut.2026.1809197

Figure Lengend Snippet: Effect of probiotic-conditioned (PP), Mycophenolic acid (MPA), and mycophenolate mofetil (MMF) on the expression of tight junction proteins. The immunoblots show the expression levels of ZO-1 (A,B) and claudin-5 (C,D) proteins in lysates of differentiated CaCo-2 cells that were exposed for 6 h to MMF or MPA (both at a concentration of 50 μM) or their vehicle, either in normal cell culture medium or PP. The bar graph shows the mean and standard deviation (SD) of the values of densitometric analysis (arbitrary unit) obtained in four experiments; symbols indicate statistical significance student’s t -test (two-group of data) analysis: * p < 0.05, ** p < 0.005 compared to untreated cells (CTRL), # p < 0.005 compared to MMF.

Article Snippet: Mycophenolate mofetil (MMF) and mycophenolic acid (MPA), produced by the Cayman Chemical Company (Ann Arbor, MI, United States), were obtained from Cabru S.A.S. (Arcore, Milan, Italy).

Techniques: Expressing, Western Blot, Concentration Assay, Cell Culture, Standard Deviation

Journal: Frontiers in Nutrition

Article Title: Enhancement of intestinal barrier function and alleviation of mycophenolic acid toxicity by a probiotic-conditioned medium in vitro

doi: 10.3389/fnut.2026.1809197

Figure Lengend Snippet: Effect of probiotic-conditioned (PP), mycophenolic acid (MPA), and mycophenolate mofetil (MMF) on cell viability in differentiated CaCo-2 cells. The cells were exposed to either MMF or MPA (both at a concentration of 50 μM) or their vehicle in normal medium or in PP for 72 h prior to measuring CyQuant fluorescence. The bar graph shows the mean and standard deviation (SD) of the values obtained in three experiments, expressed as a percentage of the control. Asterisks indicate statistical significance after the ANOVA (multiple comparison) analysis: * p < 0.05; **** p < 0.0001.

Article Snippet: Mycophenolate mofetil (MMF) and mycophenolic acid (MPA), produced by the Cayman Chemical Company (Ann Arbor, MI, United States), were obtained from Cabru S.A.S. (Arcore, Milan, Italy).

Techniques: Concentration Assay, CyQUANT Assay, Fluorescence, Standard Deviation, Control, Comparison

Journal: Frontiers in Nutrition

Article Title: Enhancement of intestinal barrier function and alleviation of mycophenolic acid toxicity by a probiotic-conditioned medium in vitro

doi: 10.3389/fnut.2026.1809197

Figure Lengend Snippet: Effect of probiotic-conditioned (PP) on reactive oxygen species (ROS) generation in differentiated CaCo-2 cells. In panel (A) cells were exposed for 2 h to mycophenolate mofetil (MMF) or mycophenolic acid (MPA) (both at a concentration of 50 μM) or their vehicle, either in normal cell culture medium or PP. In panel (B) cells were incubated in normal medium or PP for 2 h before tBHP (300 μM) was added. Intracellular ROS were measured using the dichlorofluorescein-diacetate (DCF-DA) assay after a 1 h incubation with these treatments. The bar graph shows DCF fluorescence as a percentage of control values ± SD. Asterisks indicate statistical significance after the ANOVA (multiple comparison) analysis: * p < 0.05; *** p < 0.001; **** p < 0.0001.

Article Snippet: Mycophenolate mofetil (MMF) and mycophenolic acid (MPA), produced by the Cayman Chemical Company (Ann Arbor, MI, United States), were obtained from Cabru S.A.S. (Arcore, Milan, Italy).

Techniques: Concentration Assay, Cell Culture, Incubation, Fluorescence, Control, Comparison

Journal: Frontiers in Nutrition

Article Title: Enhancement of intestinal barrier function and alleviation of mycophenolic acid toxicity by a probiotic-conditioned medium in vitro

doi: 10.3389/fnut.2026.1809197

Figure Lengend Snippet: Effect of probiotic-conditioned (PP), mycophenolic acid (MPA), and mycophenolate mofetil (MMF) on ABC1B1 activity. The bar graph shows the mean and standard deviation (SD) of Rhodamine (Rho-123) fluorescence/protein values measured in three experiments with CaCo-2 cells. The cells were incubated for 24 h with either mycophenolic mofetil (MMF) or mycophenolate acid (MPA), both at a concentration of 50 μM, in normal cell culture medium or in PP. Asterisks indicate statistical significance after the ANOVA (multiple comparison) analysis: * p < 0.05; ** p < 0.005.

Article Snippet: Mycophenolate mofetil (MMF) and mycophenolic acid (MPA), produced by the Cayman Chemical Company (Ann Arbor, MI, United States), were obtained from Cabru S.A.S. (Arcore, Milan, Italy).

Techniques: Activity Assay, Standard Deviation, Fluorescence, Incubation, Concentration Assay, Cell Culture, Comparison

Journal: Frontiers in Nutrition

Article Title: Enhancement of intestinal barrier function and alleviation of mycophenolic acid toxicity by a probiotic-conditioned medium in vitro

doi: 10.3389/fnut.2026.1809197

Figure Lengend Snippet: Transport of mycophenolic acid (MPA) across CaCo-2 transwells in the presence of probiotic-conditioned (PP) and control medium. (A) Shows representative chromatograms obtained from the upper and lower compartment medium extracts of control and PP-exposed transwells. Experiments were performed in triplicate. As detailed in the Section “2 Materials and methods,” MPA (50 μM) was added to the upper compartment of the transwell, either in normal medium (in the control group) or in the PP medium. The transwell was then incubated for 6 h before the medium was collected and processed. In all groups, MPA eluted with a retention time of approximately 18.5 min. (B) Shows bar graph of the mean ± SD of the ratios of the MPA peak areas in the upper and lower compartments of the control and PP transwells. No statistically significant difference between control and PP medium transwells was observed at the p < 0.05 significance level using an unpaired Student’s t -test.

Article Snippet: Mycophenolate mofetil (MMF) and mycophenolic acid (MPA), produced by the Cayman Chemical Company (Ann Arbor, MI, United States), were obtained from Cabru S.A.S. (Arcore, Milan, Italy).

Techniques: Control, Incubation