Cytokine production profiles of distinct ILC3 subsets in NEC mice

Experimental Design
Core Objective: Flow cytometry was performed to evaluate the functional phenotype alterations of intestinal lamina propria group 3 innate lymphoid cell (ILC3) subsets in mice with necrotizing enterocolitis (NEC), by quantifying and comparing the percentage of IL-17A-producing and IL-22-producing cells within distinct ILC3 subsets between normal control mice and NEC model mice.
Animal Model:Wild-type mice on a C57BL/6 background were used in this study. Mice in the NEC group were subjected to a standardized experimental NEC induction procedure prior to sample collection and flow cytometry detection, while mice in the control (CON) group received no NEC induction.
Experimental Groups (2 groups total, fully matched to the figure):
CON group: Normal control mice without experimental NEC induction
NEC group: Mice with standardized experimental NEC induction
Detection Targets (fully matched to the figure axes):The percentage of cytokine-producing cells within distinct intestinal lamina propria ILC3 subsets, including:
Percentage of IL-17A⁺ cells within DN ILC3s
Percentage of IL-17A⁺ cells within CCR6⁺ ILC3s
Percentage of IL-17A⁺ cells within NKp46⁺ ILC3s
Percentage of IL-22⁺ cells within DN ILC3s
Percentage of IL-22⁺ cells within CCR6⁺ ILC3s
Percentage of IL-22⁺ cells within NKp46⁺ 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.