Anti mouse cd3e fitc anti mouse cd4 pe

Flow cytometry was used to analyze the percentage of T-cell subsets CD4⁺ and CD8⁺ in the splenocytes of mice from the four groups (pVAX1-rMYR1, pVAX1, PBS, and healthy control). Splenocyte suspensions (5 × 10⁵ cells/ml) were dually stained with anti-mouse CD3e-FITC + anti-mouse CD8-PE and anti-mouse CD3e-FITC + anti-mouse CD4-PE (eBioscience, United States) for 30 min at 4°C in the dark. Cell population analysis was conducted with the FACScan flow cytometer using the CellQuest software (BD Biosciences, Franklin Lakes, NJ, United States).

The levels of CD4⁺ and CD8⁺ T cell subsets and the levels of MHC-I and MHC-II molecules in the splenocytes of mice from the four test groups, pVAX-EF-1α, pVAXI, PBS, and blank, were determined using flow cytometry as previously described [26 (link)]. Splenocyte suspensions (1 × 10⁶ cells/ml) were dually stained with anti-mouse CD3e-FITC + anti-mouse CD8-PE, anti-mouse CD3e-FITC + anti-mouse CD4-PE, anti-mouse CD3e-FITC + anti-mouse MHC-I-PE or anti-mouse CD3e-FITC + anti-mouse MHC-II-PE (eBioscience, San Diego, CA, USA) for 30 min at 4 °C in the dark. Cell population analyses were conducted with a FACScan flow cytometer with CellQuest software (BD Biosciences, Franklin Lakes, NJ, USA). Lymphocyte specific gating was set according to the forward and side scatter profiles. The percentages of CD4⁺ and CD8⁺ T lymphocytes or, MHC-I and MHC-II molecules in mouse splenocytes were determined as previously described [27 (link)].

Flow cytometry was used to detect the percentages of T cell subsets CD4+ and CD8+ in the splenocytes of mice in the pVAX1-GRA24, pVAX1, PBS, and blank groups. Splenocytes suspensions (5 ^* 10⁵ cells/ml) were dual-stained with anti-mouse CD3e-FITC+anti-mouse CD8-PE and anti-mouse CD3e-FITC+anti-mouse CD4-PE antibodies (eBioscience), for 30 min at 4°C in the dark. Cell populations were detected using a FACScan flow cytometer and analyzed using CellQuest software (BD Biosciences, Franklin Lakes, NJ, USA).