Mapk inhibitors

MAPK inhibitors were purchased from Cell Signaling Technology: 10 μM of U0126 was used to inhibit ERK1/2, 50 μM of SP600125 was used to inhibit JNK 1/2 and 10 μM of SB202190 was used to inhibit p38. In addition, 20 μM of JSH-23 (SigmaAldrich) was used to inhibit classical NFκB activation. All inhibitors were prepared as 1000x stock solutions in DMSO, thus, 0.1% DMSO was added to the control group. MEFs were exposed to the corresponding inhibitors for 30 min prior to being transferred to the scaffolds. Following 6-h incubation cells were harvested for gene expression analysis.

The DC culture medium used was Iscove's modified Dulbecco's medium (IMDM) from Gibco-BRL (Grand Island, New York, USA) containing 10% FBS from PAA Laboratories Inc. (Toronto, Canada). IL-4, IL-1β, TNFα and IFN-γ were obtained from Peprotech (Rocky Hill, New Jersey, USA). IFN-α was provided from LG Life Sciences (Chonbuk, Korea) and GM-CSF was from LG Biochemicals (Daejeon, Korea). CCL21 was purchased from R&D Systems (Minneapolis, MN, USA). Ficoll-Hypaque was purchased from Axis-SHIELD PoC AS (Lymphoprep, Oslo, Norway). All monoclonal antibodies (mAb) used for flow cytometry were obtained from BD Biosciences (Pharmingen, San Diego, CA, USA), except the mAb for CCR7 (R&D Systems, Minneapolis, MN, USA). CD14-conjugated microbeads were purchased from Miltenyi Biotec (Auburn, CA, USA). MAPK inhibitors were from Cell Signaling Technology (Boston, MA, USA) for U0126, Tocris Bioscience (Bristol, UK) for SP600125 and Calbiochem (Darmstadt, Germany) for SB203580. Wortmannin, LY294002, U73122 and BAPTA-AM were obtained from Dr Han's Laboratory (Chonnam National University, Gwangju, Republic of Korea).