Journal: Biomolecules

Article Title: Simulated Mars Gravity Impairs Intestinal Epithelial Barrier Integrity via Selective Modulation of Tight Junction Components

doi: 10.3390/biom16050739

Figure Lengend Snippet: ( a ) Scheme depicting the experimental workflow followed to grow a Caco-2 cell monolayer in simulated Mars gravity. ( b ) Representative control Caco-2 cell showing a «closed» Tight Junction (TJ) (indicated by arrowhead) by Transmission Electron Microscopy (TEM); ( c ) Representative reduced gravity-cultured Caco-2 cell showing an «open» TJ (indicated by arrowhead) by TEM; ( d ) Graph illustrating the number of open and closed TJs in control cells (white) and simulated Mars gravity (gray). Each bar represents the mean values (±SD) derived from eight independent experiments. **** = p < 0.0001, *** = p < 0.001 calculated by two-way ANOVA. ( e ) Graph illustrating the mean TEER value (±SD) measured in control (white) and simulated Mars gravity (gray) monolayers. Each bar represents the mean values (±SD) derived from three independent experiments. * = p < 0.05 calculated by unpaired t -test. Scale bars are 200 nm in ( b ) and 400 nm in ( c ).

Article Snippet: Human colon carcinoma cells (Caco-2) were obtained from ATCC (HBT-37, ATCC, Manassas, VA, USA).

Techniques: Control, Transmission Assay, Electron Microscopy, Cell Culture, Derivative Assay

Journal: Biomolecules

Article Title: Simulated Mars Gravity Impairs Intestinal Epithelial Barrier Integrity via Selective Modulation of Tight Junction Components

doi: 10.3390/biom16050739

Figure Lengend Snippet: Effect of simulated Mars gravity on cytoplasmic (cLD) and nuclear (nLD) lipid droplet content in Caco-2 cells. ( a ) Representative control Caco-2 cells without cLDs and nLDs. ( b ) Representative simulated Mars gravity-cultured Caco-2 cells showing multiple cLDs. ( c ) Representative simulated Mars gravity-cultured Caco-2 cells showing an nLD. ( d ) Graph illustrating the number of cells with nLDs on total cells counted in control (white) and simulated Mars gravity (grey) samples. Each bar represents the mean value (±SD) derived from six independent experiments. *** = p < 0.001 calculated by unpaired t -test. ( e ) Graph illustrating the percentage of the nucleus area occupied by nLDs in control cells (white) and simulated Mars gravity (grey). Each bar represents the mean value (±SD) derived from six independent experiments. *** = p < 0.001 calculated by the Mann–Whitney test. ( f ) Representative cytometry plots displaying Nile red-stained control and simulated Mars gravity-cultured Caco-2 cells. ( g ) Graph illustrating the percentage of cells exhibiting high Nile red content (within the HIGH Nile red box). Each bar represents the mean values (±SD) evaluated in four independent experiments. **** = p < 0.0001 calculated by unpaired t -test. ( h ) Representative cytometry plots displaying DCFH-DA-stained control and simulated Mars gravity-cultured Caco-2 cells. ( i ) Graph illustrating the percentage of cells within the HIGH DCFH-DA box. Each bar represents the mean values (±SD) evaluated in four independent experiments. ** = p < 0.01 calculated by unpaired t -test. Scale bar is 5 μm in ( a – c ). n, nucleus; nu, nucleolus.

Article Snippet: Human colon carcinoma cells (Caco-2) were obtained from ATCC (HBT-37, ATCC, Manassas, VA, USA).

Techniques: Control, Cell Culture, Derivative Assay, MANN-WHITNEY, Cytometry, Staining

Journal: Biomolecules

Article Title: Simulated Mars Gravity Impairs Intestinal Epithelial Barrier Integrity via Selective Modulation of Tight Junction Components

doi: 10.3390/biom16050739

Figure Lengend Snippet: ( a ) Representative Western blot of Zona occludens 1 (ZO-1), claudin-1 (CLDN1), and claudin-3 (CLDN3) in control or Mars gravity-cultured Caco-2 cells. ( b ) Graph illustrating the fold increase of densitometric analysis of ZO-1 expression levels in control cells (white) and simulated Mars gravity cells (grey). Each bar represents the mean value (±SD) derived from three independent experiments. * = p < 0.05 calculated by unpaired t -test. ( c ) Graph illustrating the fold increase of densitometric analysis of CLNDN1 expression levels in control cells (white) and simulated Mars gravity cells (grey). Each bar represents the mean value (±SD) derived from three independent experiments. ** = p < 0.01 calculated by unpaired t -test. ( d ) Graph illustrating the fold increase of densitometric analysis of CLDN3 expression levels in control cells (white) and simulated Mars gravity cells (grey). Each bar represents the mean value (±SD) derived from three independent experiments. * = p < 0.05 calculated by unpaired t -test. ( e ) Representative blot and graph illustrating the fold increase of densitometric analysis of pSTAT3/STAT3 expression levels in control cells (white) and simulated Mars gravity cells (grey). Each bar represents the mean value (±SD) derived from three independent experiments. * = p < 0.05 calculated by unpaired t -test. ( f ) Representative blot and graph illustrating the fold increase of densitometric analysis of pERK2/ERK2 expression levels in control cells (white) and simulated Mars gravity cells (grey). Each bar represents the mean value (±SD) derived from three independent experiments. p -value calculated by unpaired t -test. ( g ) Representative blot and corresponding graph illustrating the NF-κB expression level ratio between nuclear and cytoplasmic fractions in control cells (white) and simulated Mars gravity cells (grey). Each bar represents the mean value (±SD) derived from three independent experiments. p -value calculated by Unpaired t -test.

Article Snippet: Human colon carcinoma cells (Caco-2) were obtained from ATCC (HBT-37, ATCC, Manassas, VA, USA).

Techniques: Western Blot, Control, Cell Culture, Expressing, Derivative Assay

Journal: Biomolecules

Article Title: Simulated Mars Gravity Impairs Intestinal Epithelial Barrier Integrity via Selective Modulation of Tight Junction Components

doi: 10.3390/biom16050739

Figure Lengend Snippet: ( a ) Scheme depicting the experimental workflow followed to perform Calcium Switch Assay on a Caco-2 cell monolayer in simulated Mars gravity. ( b ) Representative confocal image of control and simulated Mars gravity monolayer stained for CLDN1 (green) and DAPI (blue) after Calcium Switch Assay. ( c ) Representative confocal image of control and simulated Mars gravity monolayer stained for CLDN3 (green) and DAPI (blue) after Calcium Switch Assay. ( d ) Representative confocal image of control and simulated Mars gravity monolayer stained for ZO-1 (green) and DAPI (blue) after Calcium Switch Assay. ( e ) Graph illustrating the fold increase of CLDN1 fluorescence in control cells (black) and simulated Mars gravity cells (red) with respect to T0, at different time points after Calcium Switch Assay. Each point represents the mean value (±SD) derived from three independent experiments. ** = p < 0.01 calculated by two-way ANOVA. ( f ) Graph illustrating the fold increase of CLDN3 fluorescence in control cells (black) and simulated Mars gravity cells (red) with respect to T0, at different time points after Calcium Switch Assay. Each bar represents the mean value (±SD) derived from three independent experiments. * = p < 0.05, calculated by two-way ANOVA. ( g ) Graph illustrating the fold increase of ZO-1 fluorescence in control cells (black) and simulated Mars gravity cells (red) with respect to T0, at different time points after Calcium Switch Assay. Each bar represents the mean value (±SD) derived from three independent experiments. ** = p < 0.01 calculated by two-way ANOVA. Scale bar is 10 μm in ( b – d ).

Article Snippet: Human colon carcinoma cells (Caco-2) were obtained from ATCC (HBT-37, ATCC, Manassas, VA, USA).

Techniques: Control, Staining, Fluorescence, Derivative Assay

Journal: Foods

Article Title: Valorization and Functional Enhancement of Mature Assam Tea Leaves Through Indigenous Filamentous Fungi-Based Fermentation for Functional Drink Development

doi: 10.3390/foods15091562

Figure Lengend Snippet: The cytotoxicity of Caco-2 cells ( A ) and HT-29 cells ( B ) after treatment with co-fermented mature tea leaves samples for 72 h. Different lowercase letters indicate significant differences among fermentation times within the same treatment ( p < 0.05).

Article Snippet: Human colon carcinoma cell line (Caco-2) and Human colon adenocarcinoma cell line (HT-29) obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) were used to carry out the cell cytotoxicity test.

Techniques: