Thioacetamide

All animal experiments were performed with approval from the University of Queensland Animal Ethics Committee (MED/PAH/156/13/PAHRF/NHMRC, UQDI/571/12/NHMRC/AIDRCC). To conditionally delete Wls from macrophages, homozygous Wls^flox/flox mice [40 (link)] were crossed with LysM-Cre transgenic mice [41 (link)] for 4–10 generations. Offspring with the genotype Wls^flox/flox; LysM-Cre-positive represents Wls macrophage knockouts, whilst Wls^flox/flox; LysM-Cre-negative was used as controls. Six to 9-week-old mice were administered with 30 mg/L thioacetamide (TAA, Sigma) in drinking water for 12 weeks [71 (link)] or a modified choline-deficient ethionine-supplemented (CDE) diet for 6 weeks [48 (link)], to induce chronic liver injury and fibrosis. The modified CDE diet was optimised by Professor George Yeoh [48 (link)] (UWA, Australia) and was custom made by MP Biosciences (Santa Ana, USA). The diet consisted of 70 % choline-deficient diet (Cat#0296021410) and 30 % control (choline-sufficient) diet (Cat#0296041210). At the end of the treatment period, mice were euthanised and a blood sample taken for serum isolation. Livers were perfused with 10 ml of PBS via the portal vein in situ prior to tissue harvest to minimise blood contamination. Portions of the liver were fixed in 10 % neutral buffered formalin and embedded in paraffin or homogenised in TRI reagent and stored at −80 °C for RNA isolation. The remainder of the liver was used to isolate non-parenchymal cells. Serum ALT levels were measured using the MaxDiscovery Alanine Transaminase Color Endpoint Assay (Bioo Scientific, Austin, TX, USA).

V_1.75Cr_0.25S₄ samples (VCS-450 and VCS-400) were prepared by using the hydrothermal process using a certain amount of Na₃VO₄, CrCl₃·6H₂O, thioacetamide (C₂H₅NS), and ammonium hydroxide solution dissolved in deionized water as the reactant system, which were heated at various temperatures for 12 h followed by product collection and drying (see Materials and Methods in the online Supplementary file for details).

All reagents were of analytical grade and used without further purification. Ti₃AlC₂ powder (>98 wt.% purity) was purchased from Beijing Lianli New Technology Co., Ltd., Beijing, China. By etching the Al layer of Ti₃AlC₂ with HF solution, Ti₃C₂ was obtained. In brief, 1 g Ti₃AlC₂ powder was slowly combined with 30 mL HF solution (content ≥ 40 wt.%, Xilong Chemical Co., Ltd., Shantou, China), and the mixture was stirred at room temperature for 72 h. The suspension was filtered and washed with deionized water several times, until neutral pH was achieved. The obtained Ti₃C₂ was dried under vacuum at 60 °C for 2 h.
Next, a hydrothermal reaction was employed to prepare the ZnIn₂S₄/Ti₃C₂ nanocomposites. An appropriate amount of Ti₃C₂ was dispersed in 70 mL of water by ultrasonication. To the Ti₃C₂ dispersion, ZnCl₂ (0.136 g, 1 mmol, Tianjin Fengchuan Chemical Reagent Technology Co. Ltd., Tianjing, China) and excess L-cysteine were added, and the mixture was ultrasonicated. Subsequently, InCl₃·4H₂O (0.586 g, 2 mmol., Adamas Reagent Co. Ltd., Beijing, China) and thioacetamide (TAA, 0.300 g, 4 mmol., Sinopharm Chemical Reagent Co., Ltd., Shanghai, China) were added, and the solution was transferred to a 100 mL Teflon liner, sealed in a stainless steel autoclave, and heated at 150 °C for 5 h. The product was collected by centrifugation, washed several times with de-ionized water, and dried under vacuum at 60 °C for 12 h. The mass ratio of ZnIn₂S₄ to Ti₃C₂ was varied to determine the optimal composition, and mass ratios of 2 wt.%, 5 wt.%, 10 wt.%, 20 wt.%, and 50 wt.% corresponded to samples ZIST-2, ZIST-5, ZIST-10, ZIST-20, and ZIST-50, respectively.