Intracellular fixation and permeabilization buffer set kit

Cells were stained for extracellular markers for 20 minutes at 4°C in PBS and 5% FBS. For phosphoprotein staining (PE-conjugated mouse anti–p-IRF7 antibody; catalog K47-671; BD Biosciences), cells were permeabilized and stained using the Intracellular Fixation and Permeabilization Buffer Set Kit (catalog 88-8824-00; eBioscience). Conjugated antibodies against CD123, CD303, CD15, CD14, HLA-DR, and CD10 were from BioLegend (catalog 306012, 354208, 301906, 325604, 307628, 312210, respectively). Ig isotype controls were used in all assays.

Dexamethasone (purity ≥ 98%) and OVA were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Aluminum hydroxide (AL(OH)₃) was purchased from Damao Chemical Reagent Factory (Tianjin, China). ALA (purity ≥ 90%) was purchased from Shanxi Longzhou Biological Technology Co. (Shanxi, China). TRIzol reagent was purchased from Invitrogen (Carlsbad, CA, USA). Red blood cell lysis buffer was obtained from Solarbio (Beijing, China). Mouse OVA specific IgE ELISA kit was obtained from BioLegend (San Diego, CA, USA). RPMI 1640 and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY, USA). Prime Script RT reagent kit and TB Green Premix Ex Taq II were obtained from TaKaRa (Osaka, Japan). Leukocyte activation cocktail, purified rat anti-mouse CD16/CD32 (mouse BD Fc block) antibody, and BV421-labeled anti-mouse CD3 antibody were purchased from BD (Franklin Lakes, NJ, USA). Fluorescein isothiocyanate (FITC)-labeled anti-mouse CD4 antibody, phycoerythrin (PE)-labeled anti-mouse IFN-γ antibody, allophycocyanin (APC)-labeled anti-mouse IL-4 antibody, and intracellular fixation and permeabilization buffer set kit were purchased from eBioscience (San Diego, CA, USA).