E el m0325c

Mouse blood was collected and centrifuged to extract the serum. Total IgE was measured with an ELISA kit (432401, Biolegend, USA). The lungs of mice were ground and centrifuged to extract the supernatant. The cytokines IL-4 (431104, Biolegend, USA), IL-5 (431204, Biolegend, USA), IL-13 (900-K207, PeproTech, USA), IL-33 (88-7333-88, Invitrogen, USA) and IFN-γ (430804, Biolegend, USA) in the supernatants of lung homogenates were measured using an ELISA kit. Collagen I in the lung homogenates was measured using an ELISA kit (E-EL-M0325c, Elabscience, China). Soluble CD146 in human plasma (E-EL-H2403c, Elabscience, China) was measured using commercial ELISA kits. All ELISA experiments were performed according to the instructions provided by the manufacturers.

Briefly, blood samples were kept at room temperature for 2 h and then centrifuged at 4°C (3,500 rpm, 10 min). After that, mice serum samples were collected for analysis. The serum levels of alanine transaminase (ALT) and aspartate aminotransferase (AST), which are normally used to assess liver function (Li et al., 2022b (link)), were determined according to the instructions of the ALT (C009-2–1) and AST (C010-2–1) kits (Nanjing Jiancheng Institute of Biological Engineering, Nanjing, China). To observe collagen deposition in the liver tissues, the level of hydroxyproline (Hyp) in serum was determined using hydroxyproline assay kit. The level of Col Ⅰ in serum was detected by a standard sandwich ELISA method (E-EL-M0325c, Elabscience, Wuhan, China).

To assess the total collagen level, 40 mg of the wound tissue was homogenized in 400 µL PBS and centrifuged at 12 000 g for 10 minutes at 4°C. The supernatant was harvested and subjected to the collagen I assay kit according to the manufacturer's instructions (E‐EL‐M0325c; Elabscience). The HMGB1 Enzyme‐Linked Immunosorbent Assay kit (ST51011; IBL International) was used to analyse the HMGB1 content in the human peripheral blood and drainage samples. The TGF‐β1 content in the human peripheral blood and drainage samples was measured using the TGF‐β1 Direct Enzyme‐Linked Immunosorbent Assay kit (DB100B; R&D Systems) according to the manufacturer's instructions. The absorbance was measured at 450 and 630 nm, and the concentrations were calculated according to a standard curve.