"On Day 14, 200 μg OVA were ip-injected, and then rectal temperature measured every 10 min for 70 min using a rectal thermometer (Testo, Alton, UK). After 70 min,each mouse was euthanized with CO2 inhalation, and blood obtained from the abdominal artery to permit subsequent measures of serum histamine, TNF-α, IgE, and IgG1."

Enzyme-linked immunosorbent assay (ELISA): "Levels of TNF-α, IgE, and IgG1 in RBL-2H3 cells and serum were measured by ELISA (Anlar et al. 2017). After mouse blood was clotted at room temperature, it was centrifuged at 400 g for 15 min at 4°C, and serum then obtained. RBL-2H3 cells (5 × 105 cells/well in 12-well plates) were sensitized with anti-DNP IgE (50 ng/ml). After overnight incubation, cells were treated with PFC for 30 min and challenged with DNP-HSA (1 ng/ml) for 6 hr. Each ELISA was performed in a 96-well Nunc Immuno plate using an ELISA kit (BD Biosciences, San Diego, CA) according to manufacturer’s protocols."

Statistical analysis: "All data presented in graphical format are means ± SD of independent experiments. Statistical analyses were performed using Prism5 software (GraphPad, San Diego, CA). Treatment effects were analyzed using a one-way analysis of variance (ANOVA) followed by a Dunnett’s test. A p-value < 0.05 was accepted as indicative of statistical significance."

Male ICR Mouse