Sirpα

Control, SP-R210_L(DN), and SP-R210(KO) RAW 264.7 cells were detached using non-enzymatic cell dissociation medium (Sigma-Aldrich Cat#C1544–100ML) and washed in PBS containing 2% fetal bovine serum (FBS). Cells were washed by centrifugation and discarding of supernatant, then blocked with mouse Fc block (BD Biosciences Cat#553142) in PBS at a concentration of 12.5 μg/mL and 2% FBS for 10 minutes at room temperature. After blocking, cells were washed and stained with recommended concentrations of monoclonal antibodies for 30 min at 4°C. Staining was divided into two panels; Cells were washed and placed into HBSS with 2% FBS and 0.02% sodium azide until assessment via BD LSRII flow cytometer. A minimum 30,000 events were collected and data were analyzed via FlowJo 9.8.8. Antibodies used are as follows; CD204 (Bio-Rad Cat#MCA1322A488T, 2F8); MHC II (eBioscience Cat#86–5321-42, M5/114.15.2); CD11b (BioLegend Cat#101242, M1/70); Ly6C (BD Cat#561237, AL-21); TLR2 (eBioscience Cat#12–9021-82, 6C2); F4/80 (eBioscience Cat#25–4801-82, BM8); CD36 (BD Horizon Cat#585933, CRF D-2712); SIRPα (BD Optibuild Cat#742205, P84); CD11c (eBioscience Cat#17–0114-82, N418); TLR4 (eBioscience Cat#12–9041-80, UT41); CD14 (eBioscience Cat#25–0141-82, SA2–8); SiglecF (BD Horizon Cat#562681, E50–2440)