Hmc 1

The standard reference materials (SRM) 1649b were purchased from the National Institute of Standards and Technology (NIST, Gaithersburg, MD, USA). PM particles at concentrations of 25 μg/cm² were prepared in serum-free medium.
Interleukin-1 alpha (IL-1α) and polyinosinic:polycytidylic acid sodium salt (Poly I:C), used to produce the AD-like cell model, were obtained from PeproTech (Cranbury, NJ, USA) and Sigma-Aldrich (St Louis, MO, USA). For western blotting, an anti-IL-4 antibody was obtained from Thermo Fisher Scientific (Waltham, MA, USA). An anti-IL-6 antibody was purchased from Cell Signaling (Danvers, MA, USA). An anti-IL-1β antibody was purchased from MyBioSource (San Diego, CA, USA). An anti-GAPDH antibody was obtained from Santa Cruz (Delaware Avenue, CA, USA). An anti-FLG antibody was purchased from Life Span Biosciences (Seattle, WA, USA). Anti-rabbit IgG, anti-mouse IgG, and HRP-linked antibodies were purchased from Cell Signaling Technology. The human immortalized keratinocytes (HaCaT) and human dermal fibroblast (HDF) cell lines were obtained from the Korean Cell Line Bank (Seoul, Korea). The human mast cell line HMC-1 was obtained from Merck Millipore (Temecula, CA, USA).

The following cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA): TF‐1, Kasumi‐1, NCI‐H526, NCI‐H889, NCI‐H1048, NCI‐H2170, NCI‐H446, COS7, MDA‐MB‐468, MDA‐MB‐453, MS‐1, SK‐OV‐3, CAOV‐3, and OVCAR‐3. The HMC‐1.2 and human umbilical vein endothelial cells (HUVECs) were purchased from Millipore (Burlington, MA, USA) and Lonza (Basel, Switzerland), respectively. The GIST‐T1, GIST‐430, GIST‐430/654, and GIST‐48 cells were kind gifts from S. Bauer (University Duisburg‐Essen, Germany) and J. A. Fletcher (Brigham and Women's Hospital, Boston, USA). All cells were maintained in a humidified 5% CO₂ chamber at 37 °C in the respective culture medium, following the manufacturer's instructions.

To analyze clonal growth, HMC-1 cells and HMC-1.2 (purchased from Millipore) were plated in H4230 media (StemCell Technologies, Vancouver, Canada) in the presence of kinase inhibitors. The cells were plated as duplicates or quadruplicates. HMC-1 cells with ectopic expression of TRKB were generated by retroviral transduction using a vector expressing human TRKB [12 (link)]. We performed PCR for detection of mycoplasma in cell cultures [57 (link)] and did not detect contamination of mycoplasma in tested cell lines. Primary mast cells were cultured in the presence of inhibitors for 48 hours before apoptosis analysis. Cell viability was analyzed using the Annexin-V assay (BD Pharmingen, Heidelberg, Germany). Inhibitors dasatinib and entrectinib were purchased from Selleckchem (Houston, TX). NGF and BDNF were purchased from PeproTech. We chose dasatinib for KIT inhibition in this study, since dasatinib treatment showed benefit in some patients with SM [5 (link), 58 (link)], and dasatinib is currently being tested in our Phase III clinical trial for CBF AML including KIT D816V mutated patients (ClinicalTrials.gov number, NCT02013648).