Recombinant il 9

Individuals with GD, HT, and NC donated fresh peripheral blood samples. Then PBMCs were isolated, counted (1∼2× 10⁶/ml), and plated into 6-well plates. Two separate collections of samples were generated. One group was incubated with a combination of recombinant IL-2 protein(50 ng/ml) from PeproTech (USA) and recombinant IL-9 (100 ng/ml) from R&D Systems (USA), while the other group was stimulated only with IL-2. Following a 48-hour incubation, cell supernatants were collected. The IFN-γ, IL-5, IL-13, IL-17 (IL-17A), IL-22, and IL-10 protein levels in the cell supernatant were measured using the Human High Sensitivity T Cell Magnetic Bead Panel (Merck Millipore, Germany), following the manufacturer’s protocol.

The ELISA kits of IL-4 (Cat#M4000B), IL-9 (DY409), IFN-γ (MIF00) and recombinant IL-9 were purchased from R&D Systems (Minneapolis, MN). SEB (s4881), garcinol (g5713), Annexin V kit, ChIP kit and protein G agarose beads were purchased from Sigma Aldrich (St. Louis., MO). The fluorochrome labeled antibodies of CD11c (561241), CD44 (563058), CD3 (561824), CD4 (563933), IL-9 (D9302C12), IL-4 (553047), IL-17 (560489) and IFN-γ (557998) were purchased from BD Biosciences (Franklin Lakes, NJ). The reagents for RT-qPCR and Western blotting were purchased from Invitrogen (Carlsbad, CA). Antibodies of H3K4ac, IL-9 (7923), p300 (585), pp300 (130210) and PU.1 (390405) were purchased from Biomart (Shanghai, China). The immune cell isolation kits were purchased from Miltenyi Biotech (San Diego, CA). The APAAP reagent kit was purchased from the Gukang Biotech (Guangzhou, China). The shRNA kits of VDR and p300 were purchased from Santa Cruz Biotech (Santa Cruz, CA).