Phosphatidylcholine treatment significantly downregulates IL-17A and IL-22 production in total and DN ILC3s of NEC mice

Experimental Design Core Objective: Flow cytometry was performed to evaluate the effect of phosphatidylcholine intervention on the effector cytokine production of intestinal lamina propria total group 3 innate lymphoid cells (ILC3s) and double-negative (DN) ILC3 subsets in mice with necrotizing enterocolitis (NEC), by quantifying the percentage of IL-17A-producing and IL-22-producing cells within each ILC3 subset. Animal Model:Wild-type mice on a C57BL/6 background were used in this study. All mice were subjected to standardized experimental NEC induction prior to intervention and sample collection for flow cytometry detection.
Experimental Groups (2 groups total, fully matched to the figure):
PBS group: Vehicle control group (NEC-induced mice, treated with phosphate buffered saline (PBS) vehicle) Phosphatidylcholine group: Intervention group (NEC-induced mice, treated with phosphatidylcholine)
Detection Targets (fully matched to the figure axes):The percentage of cytokine-producing cells within ILC3 subsets in the intestinal lamina propria, including: Percentage of IL-17A⁺ cells within total ILC3s Percentage of IL-17A⁺ cells within DN ILC3s Percentage of IL-22⁺ cells within total ILC3s Percentage of IL-22⁺ cells within DN ILC3s
Biological Replicates: 6 biological replicates (individual mice) per group
Statistical Analysis: Unpaired two-tailed Student's t-test was applied for comparisons between the two independent groups. Statistical significance was defined as P < 0.05.
Flow cytometry data were acquired using a CytoFLEX S flow cytometer (Beckman Coulter). The following fluorochrome-conjugated antibodies were used for cell surface and intranuclear staining in this study:NKp46-FITC, Lin-APC-Cy7, live/dead-BV510, CD4-Pacific Blue (PB), CD90.2-PE-Cy7, IL17A/IL22-PE, CCR6-APC, and RORγt-PerCP-Cy5.5.