Proportion of ILC3 subsets with pharmacological autophagy regulation

Experimental Design Core Objective: Flow cytometry was performed to evaluate the effects of pharmacological autophagy modulation (autophagy inhibition by 3-Methyladenine, autophagy activation by Rapamycin) on the proportion of total group 3 innate lymphoid cells (ILC3s) and double-negative (DN) ILC3 subsets among total innate lymphoid cells (ILCs) in the intestinal lamina propria of mice with necrotizing enterocolitis (NEC). 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 drug intervention and sample collection for flow cytometry detection. Experimental Groups (3 groups total, fully matched to the figure): DMSO group: Vehicle control group (NEC-induced mice, treated with DMSO vehicle) 3MA group: Autophagy inhibition group (NEC-induced mice, treated with 3-Methyladenine (3MA), a classical autophagy inhibitor) Rapamycin group: Autophagy activation group (NEC-induced mice, treated with Rapamycin, a classical autophagy activator)
Detection Targets (fully matched to the figure axes):The proportion of ILC3 subsets in the intestinal lamina propria among total ILCs, including: Percentage of total ILC3s among total ILCs Percentage of DN ILC3s among total ILCs
Biological Replicates: 6 biological replicates (individual mice) per group
Statistical Analysis: One-way ANOVA with Tukey’s post-hoc test was applied for multiple group comparisons. 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.