The gating strategy was shown in the previous study [23 (link)]. Doublets were removed from the analysis by setting the gate on single cells on the FSC-area (FSC-A) vs. FSC-high (FSC-H) dot plot. Cell proliferation was calculated from singlets. Next, the lymphocytes or monocytes were gated based on FSC and SSC dot plots. Then, the gate included lymphocytes, and analyses of CD4+, CD8+, and FoxP3+ cells were performed. The second sample included CD5+, CD14+, MHCII+ cells, and the median fluorescence intensity (MFI) of ROS was calculated on that cell population.
Flow cytometric analysis was performed using a FACSCanto II flow cytometer and Kaluza 1.5 software (Beckman Coulter, Brea, CA, USA); 10,000 cells of each sample were acquired. Prior to multicolor staining, the compensation was set using single-positive cells for each color.
Flow cytometric analysis was performed using a FACSCanto II flow cytometer and Kaluza 1.5 software (Beckman Coulter, Brea, CA, USA); 10,000 cells of each sample were acquired. Prior to multicolor staining, the compensation was set using single-positive cells for each color.
Free full text: Click here
