Recombinant rBmpA Protein Production

rBmpA was produced in Escherichia coli BL21 (GE Healthcare, Chicago, IL, USA) using the bacterial expression vector pGEX-6P1 (GE Healthcare) and the following primers with EcoRI and XhoI restriction sites: Forward, 5′-ACGAATTCATGAATAAAATATTGTTGTTGA-3′ and reverse, 5′-AGCTCGTAAATAAATTCTTTAAGAAA-3′ (4 (link),5 (link)). Pure ISOF was provided by Dr Weimin Yang [School of Pharmaceutical Science and Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University (KMU), Kunming, China] and reconstituted at 1×10⁵ µM/ml in dimethyl sulfoxide (DMSO) for stock solution. LPS and phorbol-12-myristate-13-acetate (PMA) were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany) and ionomycin was purchased from Apollo Scientific Ltd. (Stockport, UK). Recombinant human (rh)IL-4, rhTNF-α and rh-granulocyte macrophage colony-stimulating factor (rhGM-CSF) were purchased from R&D Systems, Inc. (Minneapolis, MN, USA).