Fig. 2: ATF3 deficiency increases macrophage/neutrophil trafficking, promotes mTOR and TLR4 activation, induces HIF-1α signaling and T cell differentiation in IR-induced liver injury.

a Liver CD11b⁺ macrophages and Ly6G⁺ neutrophils were detected by immunohistochemistry. Results were scored semi-quantitatively by averaging the number of positively stained cells (mean ± SD)/field at ×400 magnification. Representative of 4–6 mice/group. ***p < 0.001. b Quantitative RT-PCR-assisted detection of TNF-α, IL-1β, IL-6, and TGF-β expression. Mean ± SD (n = 3-4 samples/group). *p < 0.05, **p < 0.01. c Western blot analysis of phosphorylated mTOR, phosphorylated p70S6K, PHD1, HIF-1α, and TLR4. β-actin served as an internal control. Data are representative of three experiments. d, e Foxp3 and RORγt expression in spleen T cells was evaluated by flow cytometry. Representative of three separate experiments. ***p < 0.001. f ELISA analysis of serum IL-17A levels. Mean ± SD (n = 3–4 samples/group). **p < 0.01. g Cells were stained with fluorochrome-conjugated anti-F4/80 or -CD11b. F4/80 and CD11b double-positive cells were identified as infiltrating macrophages. h Western blot analysis of phosphorylated mTOR and phosphorylated p70S6K in infiltrating macrophages. β-actin served as an internal control. Data are representative of three experiments