Bv711 conjugated anti mouse cd8

Antigen (Ag)-specific CD8⁺ (Supplementary Figure 3) and CD4⁺ T cells (Supplementary Figure 5) were measured by flow cytometry using PE-and/or APC-conjugated-NP_366-374 (H-2k(b)) and PE-conjugated- NP_311-325 (I-A(b)) tetramers, respectively. Tetramers were obtained from the NIH tetramer core facility (Atlanta, GA USA). Mice were sacrificed, spleens collected and single cell suspensions were generated for each mouse as outlined above. A total of 1x10⁶ splenocytes were stained and analyzed via flow cytometry (BD bioscience LSRII). To measure the frequencies of Ag-specific CD4⁺ T cells, splenocytes were stained with the following antibodies: Live/Dead fixable aqua dead (ThermoFisher Scientic cat.no. L34957), BUV395-conjugated anti-mouse CD4 (BD cat. no. 565975), PerCP5.5-conjugated anti-mouse CD3 (Biolegend cat. no. 1002180), BV711-conjugated anti-mouse CD8 (Biolegend cat. no. 100748), PE-Cy7-conjugated anti-mouse CD44 (BD cat. no. 560569) and PE-conjugated- NP_311-325. To measure the frequencies of Ag-specific CD8⁺ cells, splenocytes were stained as follows: Live/Dead fixable aqua dead (ThermoFisher Scientic cat. no. L34957), PerCP5.5-conjugated anti-mouse CD3 (Biolegend cat. no. 1002180), BUV395-conjugated anti-mouse CD4 (BD cat. no. 565975), BV711-conjugated anti-mouse CD8 (Biolegend cat. no. 100748) and PE-Cy7-conjugated anti-mouse CD44 (BD cat. no. 560569).