The HMDM defines a toolbox composed of semi- automated DM procedures and a set of scenarios for the human to guide the analysis towards credible relations.
With the HMDM method we were able to extract relations well-established in the literature, which shows the capability of the method to find important relations in the domain.
Fourth, a computer program was developed to support the HMDM method.
Culture of THP-1 Macrophages, HMDM and COS-7 Cells THP-1 monocytes were cultured in suspension in RPMI-1640 media, supplemented with 10% heat inactivated (56[degrees]C, 30 min) fetal bovine serum (FBS), 1% penicillin/streptomycin, and 0.1% [beta]-mercaptoethanol (culture medium).
For HMDM preparation, primary human monocytes were isolated from the blood of healthy adult volunteers with ethical approval from the East London Research Ethics Committee.
Total RNA was extracted from HMDM, THP-1 macrophages, and COS-7 cells using an extraction kit (Sigma-Aldrich, Poole, UK) and RNase-free DNase (Qiagen, Crawley, UK) as recommended by the manufacturers.
For all experiments, THP-macrophages or HMDM were incubated in 24-well plates containing coverslips, which were removed for mounting at the end of the procedure.
Expression of Myo6 and Associated Proteins AP2, Dab2, and GIPC1 in THP-1 Macrophages and HMDM. To determine whether Myo6 and its binding partners are expressed in macrophages, total RNA was isolated from untreated THP-1 macrophages and HMDM and then used to generate cDNA for amplification of gene products by conventional PCR.
 demonstrated the presence of Myo6 both with and without the LI and SI in COS-7 cells, and we have used the same primer sequences to investigate their expression in THP-1 macrophages and HMDM. We found bands identical to those reported by Dance et al.
Immunoblotting indicated the presence of a protein of the molecular weight of Dab2 (96/67 kDa) in 3 lysates from THP-1 macrophages and in cells from 3 individual donors for HMDM. Images from a representative experiment are shown in Figure 3(a).