Cell culture and biological reagents

BHT101 cells, a kind gift of Prof. M. Santoro, Federico II University of Naples, were grown in Dulbecco's modified Eagle's medium (DMEM) (Sigma, St. Louis, MO USA) supplemented with 10% fetal bovine serum (Sigma). THP-1 cells were grown in RPMI 1640 medium (Sigma) supplemented with 10% fetal bovine serum (Sigma). To over-express D6/ACKR2 receptor, BHT101 cells were infected with pLenti-CMV-GFP-2A-Puro lentiviral vector (ABM, Richmond, BC, Canada) alone or containing the cDNA coding for human D6/ACKR2 fused in frame with FLAG tag. Anti-actin (sc-8432), anti-tubulin (sc-33749) and anti-IκBα (sc-371) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA), anti-FLAG (F7425 and F3165) were from Sigma, anti-D6/ACKR2 (human, PA5-11867) was from Pierce-Thermo Fisher Scientific (Waltham, MA USA), anti-F4/80 (MCA497R) was from AbD Serotec (Oxford, UK). Anti-PMCA (Plasma Membrane Calcium ATPase, clone 5F10) was from ABR Affinity BioReagents, Golden Co., USA. Anti-miR-146a was from Ambion-Thermo Fisher Scientific.