Enumeration of Inflammatory Cells in Blood

Peripheral blood samples were collected from submandibular vein and white blood cells were isolated after treatment with red blood cell lysis buffer. Cells were suspended in 100 µL of cell staining buffer (Biolegend) and incubated for 5 min with 0.5 µL of Trustain (Biolegend). Later, the cells were incubated with fluorescent conjugated antibodies at 4 °C for 45 min. For enumeration of inflammatory cells Ly6G-PE CF594 (BD Bioscience), CD11b-APC (Invitrogen), Ly6C-FITC (BD Biosciences) and F4/80-PE (Biolegend) antibodies were used. Dead cells were identified by using 7AAD (BD Pharmingen). Flow cytometry was carried out by using Accuri C6 (BD Biosciences) cytometer by following standard protocols as per the gating strategy shown in the Supplementary Fig. S1 for the enumeration of monocytes (Ly6G^-CD11b⁺Ly6C^hi), and M1 (F4/80⁺Ly6C^hi) or M2 (F4/80⁺Ly6C^lo)^{40 (link),42 (link)}.