Percentage of intestinal ILC3s, DN ILC3s, CCR6⁺ ILC3s, and NKp46⁺ ILC3s in CON and NEC groups

Experimental Design
Core Objective: Flow cytometry was performed to evaluate the alterations of group 3 innate lymphoid cell (ILC3) subsets in the intestinal lamina propria of mice with necrotizing enterocolitis (NEC), by quantifying and comparing the proportion of distinct ILC3 subsets among total innate lymphoid cells (ILCs) between normal control mice and NEC model mice.
Animal Model:Wild-type mice on C57BL/6 background were used in this study. Mice in the NEC group were subjected to standardized experimental NEC induction procedure prior to sample collection and flow cytometry detection, while mice in the control group received no NEC induction.
Experimental Groups:
CON group: Normal control mice without experimental NEC induction
NEC group: Mice with standardized experimental NEC induction
Detection Targets:The proportion of distinct ILC3 subsets among total intestinal lamina propria ILCs, including:
Percentage of double-negative ILC3s among total ILCs
Percentage of double-negative (DN) ILC3s among total ILCs
Percentage of CCR6⁺ ILC3s among total ILCs
Percentage of NKp46⁺ ILC3s among total ILCs
Biological Replicates: 7 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: Lineage-FITC, CD45-APC-Cy7, CD4-BV510, CD127-Pacific Blue (PB), CD90.2-PE-Cy7, NKp46-PE, CCR6-APC, and RORγt-PerCP-Cy5.5.