Fig. 7: Flow cytometry analysis of the impact of Rab4A and treatment by rapamycin and NAC on the surface expression of trafficked receptors in splenocyte subsets from lupus-prone mice.

B6.TC, B6.TC/Rab4A^Q72L, and B6/Rab4A^Q72L-KO mice were treated intraperitoneally (ip) three times weekly with 0.2% carboxymethylcellulose (CMC) vehicle solvent control (V), 3 mg/kg rapamycin (R), or 10 g/l of NAC (N) in drinking water. A Dimensionality reduction analyses of surface receptors by t-SNE in all mice combined. Color axis: blue (0) → red (max). Color scale from blue to red: −1622 to 262856. B Dimensionality reduction analyses by t-SNE delineated five cell clusters in response to altered expression of Rab4A and therapeutic intervention by rapamycin and NAC: cluster 1, CD71⁺CD98⁺CD152⁺ T cells; cluster 2, CD4⁺ T cells; cluster 3, CD71⁺CD98⁺SERT⁺ DN T cells; cluster 4, CD8⁺ T cells; cluster 5, CD71⁺CD98⁺CD152^- T cells. C Statistical analysis of the impact of Rab4A and treatment with rapamycin and NAC on the abundance of clusters 3, 5, and 4. Overall one-way ANOVA p values are shown in the header of each figure panel, while Sidak’s post-hoc test p values < 0.05 over brackets reflect comparison between experimental groups. The numbers (n) of mice in each experimental group were as follows: B6.TC Veh (n = 5), B6.TC Rapa (n = 5), B6.TC/Rab4A^Q72L Veh (n = 4), B6.TC/Rab4A^Q72L NAC (n = 5), B6.TC/Rab4A^Q72L Rapa (n = 6), B6.TC/Rab4A^Q72L-KO Veh (n = 3), B6.TC/Rab4A^Q72L-KO Rapa (n = 2). Charts show mean ± SEM for each experimental group.