Briefly, 106 hepatic NPCs were incubated with anti-mouse CD16/32 antibody (clone 93, Biolegend, San Diego, CA, USA) for 10 min to minimize nonspecific binding. Cells were incubated with a PerCP-conjugated anti-CD45 antibody (clone 30-F11, Biolegend), an APC-labeled anti-F4/80 antibody (clone BM8, Biolegend), a FITC-conjugated anti-CD11b antibody (clone M1/70, Biolegend), a PE-Cyanine7-conjugated anti-Ly6C antibody (clone HK1.4, Biolegend), and a PE-conjugated anti-CCR2 antibody (clone SA203G11, Biolegend) or incubated with a PerCP-conjugated anti-CD45 antibody (clone 30-F11, Biolegend), an APC-conjugated anti-CD3 antibody (clone 17A2, Biolegend), a PE-Cyanine7-conjugated anti-NK1.1 (clone PK136, Biolegend), a PE-conjugated anti-CD4 (clone GK1.5, Biolegend), and a FITC-conjugated anti-CD8a (clone 53-6.7, Biolegend). The results were collected by a BD FACSAria III Flow Cytometer (BD Bioscience, San Jose, CA, USA) and analyzed using FlowJo 10.4 software (BD Bioscience).
After the cells were treated with hypoxia-reoxygenation (H/R), apoptosis was assessed by an Annexin V-FITC/PI Apoptosis Detection Kit (Vazyme Biotech, Nanjing, China) based on the manufacturer’s protocols.
After the cells were treated with hypoxia-reoxygenation (H/R), apoptosis was assessed by an Annexin V-FITC/PI Apoptosis Detection Kit (Vazyme Biotech, Nanjing, China) based on the manufacturer’s protocols.