Fig. 7.

OASL1 is essential for inhibiting G3BP1/TBK1 activation and cell death in myeloid TXNIP-deficient livers in response to IR.

The TXNIP^M-KO mice were injected via tail vein with OASL1 siRNA (2.5 mg/kg) or NS control siRNA mixed with mannose-conjugated polymers at 4 h before ischaemia. (A) Immunofluorescence staining of AlexaFluor488-labelled control siRNA and CD68-positive macrophages in ischaemic liver tissue (n = 6 mice/group), Scale bars, 25 μm. (B) Representative histological staining (H&E) of ischaemic liver tissue (n = 6 mice/group) and Suzuki’s histological score. Scale bars, 200 μm. (C) Immunofluorescence staining of CD11b⁺ macrophages in ischaemic livers (n = 6 mice/group). Quantification of CD11b⁺ macrophages. Scale bars, 100 μm. (D) Immunohistochemistry staining of Ly6G⁺ neutrophils in ischaemic livers (n = 6 mice/group). Quantification of Ly6G⁺ neutrophils. Scale bars, 200 and 50 μm. (E) Western blot analysis and relative density ratio of G3BP1, p-TBK1, and p-IRF3. Representative of 4 experiments. (F) Liver apoptosis by TUNEL staining in ischaemic livers (n = 6 mice/group). Results were scored semiquantitatively by averaging the number of apoptotic cells. Scale bars, 100 μm. All data represent the mean ± SD. Statistical analysis was performed using the permutation t test. ∗∗p <0.01, ∗∗∗p <0.005. G3BP1, Ras GTPase-activating protein-binding protein 1; HPF, high power field; IR, ischaemia/reperfusion; IRF3, interferon regulatory factor 3; NS, non-specific; OASL1, 2′,5′ oligoadenylate synthetase-like 1; siRNA, small interfering RNA; TBK1, TANK-binding kinase 1; TXNIP, thioredoxin-interacting protein; TXNIP^M-KO, myeloid-specific TXNIP knockout.