Pe conjugated anti mouse ly6g clone rb6 8c5

Mouse primary liver cells were isolated following a previously established method (You et al. 2008 (link)). In brief, the mouse liver was perfused with Hank's balanced salt solution followed by a digestion buffer containing 0.2% collagenase (Type IV, Sigma). Single cell suspensions were filtered through a 100 µm cell strainer and centrifuged at 50 g for 3 min. Hepatocytes were in the pellet, and the cells in the supernatant were collected and further centrifuged in 35% percoll (Sigma, Germany) to obtain liver NPCs. Red blood cells were lysed with red blood cell lysing buffer. Liver NPCs were stained with fluorochrome-conjugated antibodies for multi-color fluorescent-activated cell sorting (FACS) analysis, including APC Cyanine7conjugated anti-mouse CD45 (clone 30F11, #130-105-506; Miltenyi Research Inc. San Diego, CA), APC conjugated anti-mouse F4/80 (clone BM8, #17-4801-82, eBioscience), PE-vio770 conjugate anti-mouse CD11b (clone M1/70, #25-0112-82, eBioscience), PE-conjugated anti-mouse Ly6G (clone RB6-8C5, #561084, BD Biosciences) and FITC-conjugated anti-mouse Ly6C (clone AL21, #553104, BD Biosciences, San Jose, CA). BD FACSCanto II flow cytometer was used and the software FlowJo (V10) was utilized to analyze data.

Freshly isolated LNPCs were incubated with mouse Fc receptor blocker to prevent non-specific binding. Then, the cells were incubated with various staining antibodies, including APC Cyanine7conjugated anti-mouse CD45 (clone 30F11, #130-105-506; Miltenyi Research Inc. San Diego, CA), PE-vio770 conjugate anti-mouse CD11b (clone M1/70, #25-0112-82, eBioscience), APC conjugated anti-mouse F4/80 (clone BM8, #17-4801-82, eBioscience), FITC-conjugated anti-mouse Ly6C (clone AL21, #553104, BD Biosciences, San Jose, CA) or PE-conjugated anti-mouse Ly6G (clone RB6-8C5, #561084, BD Biosciences).