Hdac11

The LPS, FITC-labeled dextran, calcitriol and ChIP kit were purchased from Sigma Aldrich (St. Louis., MO). The antibodies of VDR, ZO-1, claudin-5, occludin, HDAC11, acH3, acH4, shRNA kits of VDR and HDAC11 were purchased from Santa Cruz Biotech (Santa Cruz, CA). The reagents for Western blotting and RT-qPCR were purchased from Invitrogen (Carlsbad, CA). The reagents used in this study contained <0.2U endotoxin/10 μg reagents as assessed using the Limulus assay (Limulus amebocyte lysate QCL 1000, Bio Whittaker, Walkersville, MD, USA).

ChIP assay was performed with DCs using a reagent kit (Sigma Aldrich) following the manufacturer's instructions. Briefly, DCs were fixed with 1% formaldehyde for 15 min, lysed and sonicated to shear the chromatin DNA to 100–500 bp. Cell lysates were precleared by incubation with protein G-agarose beads for 2 h at 4°C. The supernatant was collected and incubated overnight at 4°C with 2 μg of antibodies of HDAC11 or c-Maf (Santa Cruz Biotech) or isotype IgG (a negative control). The antibody-chromatin complex was precipitated by incubation with protein G-agarose beads for 1 h at 4°C. The beads were centrifuged, washed and eluted in elution buffer. DNA was recovered from the precipitated samples by reverse crosslinking at 65°C for 4 h and digested with proteinase K for 1 h at 45°C to remove proteins, then the immunoprecipitated DNA was recovered by phenol/chloroform extraction and ethanol precipitation. The DNA or input was analyzed by PCR with the miR-19a promoter primers (Beijing Yijie Biotech; Beijing, China). The results were presented as folds of input.

The fluorochrome-labeled antibodies for flow cytometry were purchased from eBioscience (San Diego, CA). The antibodies of IL-10, A20, HDAC11, Sp, IRF, AP, CREB, c/EBP, c-Maf, NF-κB, STAT3, HDAC11 shRNA kit and A20 shRNA kit were purchased from Santa Cruz Biotech (Santa Cruz, CA). LPS, ChIP kit and luciferase kit were purchased from Sigma Aldrich (St. Louis., MO). The recombinant IL-4, IL-5, IL-13 and the ELISA kit of IL-4 were purchased from R&D Systems (Minneapolis, MN). The OVA-specific IgE ELISA kit was purchased from the Biomart (Beijing, China). The reagent kits for magnetic cell sorting were purchased from Miltenyi Biotech (San Diego, CA). Reagents for RT-qPCR and Western blotting were purchased from Invitrogen (Carlsbad, CA).