Figure 1.

Single-cell RNAseq analysis of tetramer-positive T cells in Blastomyces-challenged mice following subcutaneous (SC) or intranasal (IN) vaccination

(A)Experimental schema for IN and SC vaccination with Blastomyces endoglucanase-2 (Bl-Eng2), lethal experimental challenge with Blastomyces, sorting of Bl-Eng2-specific CD4⁺T cells with tetramer, and single-cell RNAseq.

(B) Gating strategy for selection of tetramer⁺ CD4⁺T cells. Representative flow cytometry plots shown for the SC group cells; see Figure S1 for IN group flow cytometry plots.

(C) Uniform manifold approximation and projection (UMAP) for integrated analysis of SC and IN group cells yields 16 distinct cell clusters. Cluster numbers are assigned based on largest population (cluster 1) to smallest (cluster 16).