Figure 3. aCD3-LNPs transfect splenic and circulating CD4⁺/CD8a⁺ T cells.

A-D) mCherry⁺ CD4⁺ (A,C) or mCherry⁺ CD8a⁺ (B, D) T cells as a percentage of CD4⁺ or CD8a⁺ T cells, respectively, at 24 and 48h after control (no treatment, isotype conjugated LNPs, or aCD3 F(ab’)₂) or 1, 2, or 16% aCD3-LNP treatment in the spleen (A, B) or blood (C, D) (n=28). E-F) Normalized count of mCherry⁺ CD3e⁺, CD4⁺, and CD8a⁺ T cells at 24 h after 16% aCD3-LNP treatment from spleen (E) or blood (F). Each replicate was normalized in its respective 1, 2, or 16% group using the following: Normalized Count of Analyzed mCherry⁺ T cells = [(largest T-cell count among replicates in the group) / (T-cell count of replicate)] · mCherry⁺ count of replicate. All data are plotted as mean ± SD. For plots with more than 2 groups, statistical analyses were performed using ordinary one-way ANOVAs with Tukey’s multiple comparison test. Plots with 2 groups were statistically analyzed using an unpaired T test with Welch’s correction, ns: not significant. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001. ψ represents p<0.0001 of the below group across all other groups unless otherwise denoted with statistical analysis lines above.