RAW264.7 cells group as mentioned earlier. The apoptosis detection kit (556547, Becton Dickinson Company, United States) was used following manufacturer’s instructions. Shortly, cells at suitable concentrations were incubated with 5 μl FITC Annexin V and 5 μl PI for 15 min in dark. Then, we analyzed apoptosis by FACS within 1 h. Cells that were stained with FITC Annexin V (+) and PI(+) or (-) were considered apoptotic.
Cells were washed using 3 ml 1% BSA-PBS. Then, we suspended them in 1% BSA-PBS and counted their number with a cell counter, followed by pre-incubation with mouse TruStain FcX PLUS (anti-mouse CD16/32)(Biolegend, 156604, CA, USA)for 10 min. The cell suspension was stained with anti-mouse F4/80 BV605 (743281, Becton Dickinson, San Diego, CA, United States), APC anti-mouse CD163 (155,306, Biolegend, CA, United States), and anti-mouse CD86 BV421(105031, Biolegend, CA, United States) at 4°C for 30 min in the dark. After washing and resuspending in PBS, the cells were analyzed by FACS. The gating strategy was listed in (Supplementary Figure S1 ).
Cells were washed using 3 ml 1% BSA-PBS. Then, we suspended them in 1% BSA-PBS and counted their number with a cell counter, followed by pre-incubation with mouse TruStain FcX PLUS (anti-mouse CD16/32)(Biolegend, 156604, CA, USA)for 10 min. The cell suspension was stained with anti-mouse F4/80 BV605 (743281, Becton Dickinson, San Diego, CA, United States), APC anti-mouse CD163 (155,306, Biolegend, CA, United States), and anti-mouse CD86 BV421(105031, Biolegend, CA, United States) at 4°C for 30 min in the dark. After washing and resuspending in PBS, the cells were analyzed by FACS. The gating strategy was listed in (