Genetically Modified Tim-4 Mutants

All plasmids in this study were constructed and sequenced to confirm their identity. The Tim-4, Tim-4^AAA, Tim-4^TL, Tim-4-GPI, and Anxa5-GPI plasmid used in this study have been previously described^{33 (link)}. Tim-4^ΔIgV lacking the IgV domain of Tim-4 (residues 25–134) and Tim-4^ΔMucin without the mucin domain (residues 135–270) were constructed into the pDisplay vector using a PCR-based method. The antibodies used in the study were anti-HA antibody (Santa Cruz, SC-7392), PE-conjugated anti-F4/80 antibody (Biolegend, 123110), Alexa Fluor 405-conjugated anti-mouse antibody (Invitrogen, A31553), Alexa Fluor 488-conjugated anti-mouse antibody (Invitrogen, A-11029), and Alexa Fluor 555-conjugated anti-mouse antibody (Invitrogen, A-21422). Polystyrene beads (FluoSpheres carboxylate beads, Invitrogen, F8826), FITC-labeled E. coli (E-2861), S. aureus (S-2851), and zymosan A (Z-2841) bioparticles were purchased from Invitrogen. Alexa Fluor 488-conjugated LPS (L-23351) was purchased from Molecular Probes. LPS from E. coli O55:B5 (L2880) was purchased from Sigma Aldrich. About 6.0–8.0-μm streptavidin-coated polystyrene particles (SVP-60-5) were purchased from Spherotech. For IgG opsonization, FITC-conjugated streptavidin antibody (200-402-095) was purchased from Rockland Immunochemicals. Dexamethasone (265005) and TAMRA-SE (C1171) were purchased from Merck and ThermoFisher, respectively.