Fig. 5: Requirement of TRMT61A for ILC3-Mediated protection against Citrobacter rodentium infection.

a Wild-type mice were orally inoculated with 1 × 10⁹ Citrobacter rodentium or PBS as a control. Three days post-infection, intestinal LPLs were isolated. ILC3s (Lin^–CD45.2^intCD90.2^hi) were FACS sorted, and mRNA was extracted. Real-time RT-PCR was conducted to analyze the expression of Trmt61a in ILC3s from infected and control mice. n = 5–7 mice per group. b–n Trmt61a^fl/fl and Trmt61a^ΔRorc mice were infected with 1 × 10⁹ Citrobacter rodentium. Flow cytometry analysis 5 days post-infection: b, e, h, k, m Representative flow cytometry plots showing various immune cell populations. Population frequencies of total ILC3s (c), IL-22⁺ ILC3s (f) and IL-17⁺ ILC3s (i). Counts of total ILC3s (d), IL-22⁺ ILC3s (g) and IL-17⁺ ILC3s (j). l Population frequency of Ki67⁺ ILC3s, indicating proliferation. n Population frequency of total CD4⁺ T cells. Data from day 5 post-infection, n = 7 mice per group. o–r Morphological and Histological Assessments: changes in body weight (o), representative image of the large intestine (p), colon length measurements (q), histology scores assessing tissue damage (r). Data from day 5 post-infection, n = 8 mice per group. s Bacterial Load Analysis: quantification of bacterial load in feces on days 3 and 5 post-infection. Data are pooled from two independent experiments, presented as means ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.