Percentage of CD4⁻RORγt⁺ and CD4⁺RORγt⁺ cells among total CD45⁺ cells

Experimental Design
Core Objective: Flow cytometry was performed to evaluate the alterations of RORγt-expressing immune cell subsets in the intestinal lamina propria of mice with necrotizing enterocolitis (NEC), by quantifying and comparing the proportion of CD4⁻RORγt⁺ group 3 innate lymphoid cells (ILC3s) and CD4⁺RORγt⁺ T helper 17 (Th17) cells among total CD45⁺ immune cells 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 proportion of target RORγt⁺ immune cell subsets among total CD45⁺ immune cells in the intestinal lamina propria, including:
Percentage of CD4⁻RORγt⁺ ILC3s among CD45⁺ cells
Percentage of CD4⁺RORγt⁺ Th17 cells among CD45⁺ cells
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.
The following fluorochrome-conjugated antibodies were used for cell surface and intranuclear staining in this study: CD45-APC-Cy7, CD4-BV510 and RORγt-PerCP-Cy5.5.