Ab108870

The whole blood samples of four mice in each group were centrifuged at 3500 rpm and 4°C for 15 min to obtain serum samples. Content of serum IL-2 (ab10752, Abcam, U.S.A.), IL-10 (ab108870, Abcam, U.S.A.) and TNF-α (ab6671, Abcam, U.S.A.) were measured according to the instructions of the kits.

The preserved serum of mice in each group was centrifuged at 1,509.3 × g for 15 min, and the contents of immune factors TNF-α, IL-6, IL-10, and TGF-β were detected in strict accordance with the operating steps of the ELISA antibodies: TNF-α (ab208348, Abcam, Cambridge, UK), IL-6 (ab222503, Abcam, Cambridge, UK), IL-10 (ab108870, Abcam, Cambridge, UK), and TGF-β (ab119557, Abcam, Cambridge, UK).

Two weeks after surgery, the concentration of IL-10 (ab108870; Abcam) in the muscle was assessed using commercially available ELISA kits (USCN Business Co., Ltd.) according to the manufacturer's protocol.

The supernatant of the spleen homogenate was harvested in order to determine the levels of cytokines as per the protocol of the IL-10 (ab108870, Abcam Inc.) and IL-4 (ab100710, Abcam Inc.) ELISA kit. The optical density (OD) value at 450 nm was measured using a totipotent enzyme marker (Synergy 2, Winooski, VT, USA). The standard concentration was utilized as the X-axis and OD value as Y-axis in order to determine the regression equation of the standard curve. The OD value was substituted into the equation to calculate the target protein concentration in the samples.

In cultured cells, GFP was detected by direct fluorescence. The mouse spine tissue was fixed with formalin and embedded in paraffin. Thin sections of the tissue were then cut using a microtome and placed onto glass slides. The sections are then deparaffinized and rehydrated using a series of alcohol solutions. The sections are then stained with hematoxylin, which stains the nuclei of the cells blue, and eosin, which stains the cytoplasm and extracellular matrix pink. The slides are then dehydrated again and coverslipped. Enzyme-linked immunosorbent assay (ELISA) was performed to measure the levels of mouse Ccl2 (ab208979; Abcam, Hangzhou, China), mouse Il1β (ab197742; Abcam), mouse tumor necrosis factor alpha (TNFα, ab208348; Abcam), mouse interferon gamma (IFNɣ, ab282874; Abcam), mouse Il6 (ab100712, Abcam), mouse Il17 (ab100702; Abcam), mouse arginase 1 (ARG1, ab269541; Abcam), mouse CD163 (ab272204, Abcam), mouse transforming growth factor β1 (TGFβ1, ab119557, Abcam) and mouse Il10 (ab108870, Abcam) in cell lysis. Specific kits were utilized as per the manufacturer’s instructions.