Single cell suspensions were obtained from uninfected skin and lymph nodes as previously described [13] (link). Intracellular CD207 staining was performed using BD Bioscience Cytofix/Cytoperm kit (BD Biosciences, San Jose, CA). For extracellular staining, antibodies were diluted in staining media (3% Calf Serum, 5mM EDTA, 0.04% Azide) with Fb block. The following antibodies were used for skin: CD45.2 AF488, CD64 PE, TCRβ PE-Dazzle 594, CD3ε PE-Cy7, Gr1 AF 647, MHCII AF 700, CD11b BV421, TCRγδ BV510 (all Biolegend, SanDiego, CA), CD11c PerCPCy5.5 (Tonbo Biosciences, San Diego, CA), CD90.2 BUV395, CD8a BUV737 (both BD Biosciences, San Jose, CA). For lymph node staining we used: CD45.2 AF488, CD64 PE, Gr1 AF 647, MHCII AF 700, CD11b BV421, (all Biolegend, SanDiego, CA), CD11c PerCPCy5.5 (Tonbo Biosciences, San Diego, CA). Sample data was acquired on an LSRFortessa flow cytometer (Becton Dickinson, Franklin Lakes, NJ), and analysed using FlowJo software (TreeStar, Ashland, OR).
Edwards T.N., Zhang S., Liu A., Cohen J., Zhou P.Y., Mogavero S., Hube B., Berman J., Bougnoux M., Mathers A.R., Gaffen S.L., Albers K.M., Koerber H.R., Davis B.M., d’Enfert C., & Kaplan D.H. (2020). Extracellular ATP released from Candida albicans activates non-peptidergic neurons to augment host defense.
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