Journal: Journal of dairy science

Article Title: Milk osteopontin has high iron-binding capacity and facilitates iron absorption in intestinal cells.

doi: 10.3168/jds.2024-25305

Figure Lengend Snippet: Figure 7. Ferritin content of Caco-2 cells in the presence of phytic acid. Caco-2 monolayers were incubated for 3 h with FeSO4 (A), OPN-Fe (B), or dephosphorylated OPN-Fe (deOPN-Fe; C) corresponding to 20 µM iron in the presence or absence of 200 µM phytic acid. Intracellular ferritin levels were quantified by ELISA and normalized to total cell protein. Data are presented as mean ± SD (n = 3) and are representative of 3 individual experiments. An unpaired 2-tailed t-test was performed, and different lowercase letters (a,b) indicate significant differences (P < 0.001 [A], P < 0.01 [C]).

Article Snippet: The cell ferritin levels were analyzed using a Human Ferritin DuoSet ELISA (R&D Systems) following the manufacturer’s protocol.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Journal of dairy science

Article Title: Milk osteopontin has high iron-binding capacity and facilitates iron absorption in intestinal cells.

doi: 10.3168/jds.2024-25305

Figure Lengend Snippet: Figure 6. Viability, ferritin content, and expression of DMT1 of Caco-2 cells treated with iron or OPN-Fe. Caco-2 monolayers were incubated with FeSO4 or OPN-Fe subjected to gastric (G) or gastrointestinal (GI) digestion for 3 h (20 µM iron in each experiment). (A) Cytotoxicity of OPN- Fe complexes was evaluated using an MTT cell viability assay. (B) Intracellular ferritin levels were quantified by ELISA and normalized to total cell protein. (C) DMT1 mRNA levels were quantified by real-time quantitative PCR and normalized to the endogenous control β-actin. Data are presented as mean ± SD (n = 8 [A] or n = 3 [B, C]) and are representative of 2 (A, C) or 3 (B) individual experiments. One-way ANOVA followed by Tukey’s multiple comparisons test (A, B) or an unpaired 2-tailed t-test (C) was performed, and different lowercase letters (a,b) indicate significant differences (P < 0.0001 [A], P < 0.001 [B], P < 0.05 [C]).

Article Snippet: The cell ferritin levels were analyzed using a Human Ferritin DuoSet ELISA (R&D Systems) following the manufacturer’s protocol.

Techniques: Expressing, Incubation, Viability Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Control