Recombinant il 4

Embryonic thymus lobes at E15 (d 15) of gestation were placed in 1.35 mM 2dGuo, as previously described (Cowan et al., 2013 (link)). 2dGuo FTOC were then either transplanted in vivo or were used for in vitro experiments involving a further 4-d stimulation in the presence or absence of 100 µg/ml recombinant IL-4 (BioLegend) and IL-13 (PeproTech).

Acetone, 3-mercaptopropionic acid, DSPE and PEG₂₀₀₀ were obtained from RuixiBio (Xi’an, China). DMF and PFOB were purchased from Sigma-Aldrich (St. Louis, USA). Antibodies against Nrf2, collagen Ia1, α-smooth muscle actin (SMA), F4/80, and CD206 were purchased from Cell Signaling Technology (MA, USA). CD86 was purchased from Invitrogen (Carlsbad, CA, USA). Antibodies against GAPDH and HO-1 were purchased from Proteintech (Hubei, China). The HR-conjugated goat anti-rabbit secondary antibody was obtained from Abbkine Scientific (Wuhan, China). Recombinant IL-4 was purchased from BioLegend (CA, USA). The ELISA kits for TGF-β, IL-4, and IL-13 were purchased from Multisciences (Lianke) Biotech (Hangzhou, China), and the test kits for superoxide dismutase (SOD) and malondialdehyde (MDA) were purchased from Beyotime Biotechnology Company (Shanghai, China). DiR was purchased from Bioss (Beijing, China), and DiI and 4',6-diamidino-2-phenylindole (DAPI) were purchased from Beyotime Biotechnology Company (Shanghai, China). Other reagents used in western blot analysis were purchased from Solarbio (Beijing, China).

Starting from D8 after tumor inoculation, each mouse was injected subcutaneously 4 times at the inguinal area with 25 μg of recombinant IL-4, IL-6, or IL-21 (BioLegend, USA) daily (D8, D9, D10, and D11). In parallel, mice injected with PBS in the same way were set as control.