Journal: Biology

Article Title: Selective Targeting of Senescent FHs74Int Cells by Human Breast Milk Free Fatty Acids

doi: 10.3390/biology14101355

Figure Lengend Snippet: Detection of senescent cells in FHs74 Int cells. ( A ) Representative β-galactosidase staining of FHs74 cells showing the presence of senescent cells. ( B ) Representative histogram of the expression of p16INK4a (left) and p21 (right) assessed by flow cytometry. Data are presented as Mean ± standard error of the mean (SEM), n = 4. ( C ) Representative flow cytometry dot plot image of Pyronin Y/7-AAD dual staining of freshly thawed FHs74.

Article Snippet: The FHs74 Int cells (FHs74) were obtained from the American Type Culture Collection (ATCC; CCL-241; Manassas, VA, USA).

Techniques: Staining, Expressing, Flow Cytometry

Journal: Biology

Article Title: Selective Targeting of Senescent FHs74Int Cells by Human Breast Milk Free Fatty Acids

doi: 10.3390/biology14101355

Figure Lengend Snippet: Impact of HBM on FHs74 Int cells. ( A ) Representative β-galactosidase staining of FHs74. cells exposed or not to NonCyt or Cyt-HBM. ( B ) Flow cytometry cell counts of viable (7-AAD negative) FHs74 cells recovered after exposure to different HBM samples. ( C ) Flow cytometry expression of Annexin V. Data are presented as Mean ± standard error of the mean (SEM); * p ≤ 0.05, ** p ≤ 0.01. n = 4–5 different HBM samples. Note: The difference in sample size between the FHs74-only condition (n = 4) and the HBM-treated conditions (n = 5) is due to the experimental design, in which multiple HBM donors were tested simultaneously against a single FHs74Int control within the same experiment. FHs74 = FHs74 intestinal cells alone; HBM = Human breast milk; FHs74 + FM = FHs74 intestinal cells exposed to formula milk; FHs74 + HBM = FHs74 intestinal cells exposed to human breast milk.

Article Snippet: The FHs74 Int cells (FHs74) were obtained from the American Type Culture Collection (ATCC; CCL-241; Manassas, VA, USA).

Techniques: Staining, Flow Cytometry, Expressing, Control

Journal: Biology

Article Title: Selective Targeting of Senescent FHs74Int Cells by Human Breast Milk Free Fatty Acids

doi: 10.3390/biology14101355

Figure Lengend Snippet: Quantification of β-galactosidase activity in FHs74 cells. ( A ) Luminescence signal measured by the beta-glo assay following exposure to different HBM samples. ( B ) Two categories of HBM based on their β-galactosidase-reducing activity. Data are presented as Mean and standard error of the mean (SEM); ** p ≤ 0.01; **** p ≤ 0.0001. n = 14 different donors of HBM. Note: Quantification of β-galactosidase activity in FHs74Int cells exposed to different HBM samples. Error bars are not visible for the FHs74Int and Noncyt-HBM conditions due to the absence of variation between samples (100% signal in both cases). FHs74 = FHs74 intestinal cells alone; HBM = Human breast milk; FHs74 + HBM = FHs74 intestinal cells exposed to human breast milk.

Article Snippet: The FHs74 Int cells (FHs74) were obtained from the American Type Culture Collection (ATCC; CCL-241; Manassas, VA, USA).

Techniques: Activity Assay, Glo Assay

Journal: Biology

Article Title: Selective Targeting of Senescent FHs74Int Cells by Human Breast Milk Free Fatty Acids

doi: 10.3390/biology14101355

Figure Lengend Snippet: Expression of signaling molecules after HBM exposure. Expression of phosphorylated p38 ( A ) and NFκB ( B ) in FHs74 cells following treatment with Cyt- or Noncyt-HBM. Data are presented as Mean and standard error of the mean (SEM); **** p ≤ 0.0001. n = 5–9 different donors of human breast milk. Note: The difference in sample size between the FHs74-only condition (n = 4–5) and the HBM-treated conditions (n = 5–9) is due to the experimental design, in which multiple HBM donors were tested simultaneously against a single FHs74Int control within the same experiment . FHs74 = FHs74 intestinal cells alone; FHs74 + FM = FHs74 intestinal cells exposed to formula milk; FHs74 + HBM = FHs74 intestinal cells exposed to human breast milk.

Article Snippet: The FHs74 Int cells (FHs74) were obtained from the American Type Culture Collection (ATCC; CCL-241; Manassas, VA, USA).

Techniques: Expressing, Control

Journal: Biology

Article Title: Selective Targeting of Senescent FHs74Int Cells by Human Breast Milk Free Fatty Acids

doi: 10.3390/biology14101355

Figure Lengend Snippet: Cellular stress and apoptosis markers. ( A ) Caspase 3/7 activity in FHs74 cells exposed to Cyt- and Noncyt-HBM. ( B ) γH2AX expression levels indicate DNA damage. ( C ) Mitochondrial membrane potential measured by DiOC 2 (3) staining. Data are presented as Mean and standard error of the mean (SEM); * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001. n = 5 different donors of human breast milk. Note: The difference in sample size between the FHs74-only condition (n = 4) and the HBM-treated conditions (n = 5) is due to the experimental design, in which multiple HBM donors were tested simultaneously against a single FHs74Int control within the same experiment. FHs74 = FHs74 intestinal cells alone; FHs74 + FM = FHs74 intestinal cells exposed to formula milk; FHs74 + HBM = FHs74 intestinal cells exposed to human breast milk.

Article Snippet: The FHs74 Int cells (FHs74) were obtained from the American Type Culture Collection (ATCC; CCL-241; Manassas, VA, USA).

Techniques: Activity Assay, Expressing, Membrane, Staining, Control

Journal: Biology

Article Title: Selective Targeting of Senescent FHs74Int Cells by Human Breast Milk Free Fatty Acids

doi: 10.3390/biology14101355

Figure Lengend Snippet: Fatty acid fraction and HBM cytotoxicity. ( A ) Quantification of FFA levels across HBM samples ( n = 10). ( B ) Correlation between FFA concentration and Beta-glo signal ( n = 10). ( C ) Beta-glo signal after exposure to whole HBM, aqueous, and fatty fractions ( n = 4). Data are presented as Mean and standard error of the mean (SEM); ** p ≤ 0.01; **** p ≤ 0.0001. FHs74 = FHs74 intestinal cells alone; FHs74 + FM = FHs74 intestinal cells exposed to formula milk; FHs74 + HBM = FHs74 intestinal cells exposed to human breast milk.

Article Snippet: The FHs74 Int cells (FHs74) were obtained from the American Type Culture Collection (ATCC; CCL-241; Manassas, VA, USA).

Techniques: Concentration Assay

Journal: Biology

Article Title: Selective Targeting of Senescent FHs74Int Cells by Human Breast Milk Free Fatty Acids

doi: 10.3390/biology14101355

Figure Lengend Snippet: FFA concentration and β-galactosidase signal. ( A ) Quantification of FFA levels in Noncyt-HBM samples prior and after 4 days at 37 °C. ( B ) Beta-glo signal of FHs74 intestinal cells to Noncyt-HBM samples prior and after 4 days at 37 °C. Data are presented as Mean and standard error of the mean (SEM); * p ≤ 0.05; *** p ≤ 0.001. n = 4 different donors of human breast milk. HBM = human breast milk; FFA = Free fatty acids.

Article Snippet: The FHs74 Int cells (FHs74) were obtained from the American Type Culture Collection (ATCC; CCL-241; Manassas, VA, USA).

Techniques: Concentration Assay