(A and B) Flow cytometric analysis of key signaling molecules in mTORC1 signaling pathways in CD4⁺GZMB⁺ cells and CD4⁺GZMB^– cells from patients with GO (n = 4). FACS plots and bar plots were presented for intracellular p-mTOR (A) and its downstream molecule p-S6K (B). CD4⁺GZMB⁺ cells expressed higher levels of p-mTOR and p-S6K than CD4⁺GZMB^– cells. (C) Representative images of Western blot, which analyzed the key signaling molecules involved in mTORC1 signaling pathways from PBMCs treated with or without rapamycin. The PBMCs were obtained from patients with GO and incubated with or without rapamycin (100 nM, 48 hours). (D) Bar plots for relative intensities in the Western blot of p-mTOR and p-S6K in PBMCs treated with or without rapamycin (n = 3). (E) Flow cytometric analysis of the proportions of CD4⁺ CTLs expressing GZMB in PBMCs treated with or without rapamycin (n = 3). FACS plots and bar plots are presented. (F) Flow cytometric analysis of CCL5 in CD4⁺GZMB⁺ cells treated with or without rapamycin (n = 3). FACS plots and bar plots are presented. (G and H) Flow cytometric analysis of the proportions of CD4⁺PRF1⁺ and CD4⁺IFNG⁺ cells in PBMCs treated with or without rapamycin (n = 3). FACS plots and bar plots are presented. Values represent the mean ± SEM. *P < 0.05 and **P < 0.01, by 2-tailed, paired samples t tests for A and B and 2-tailed, independent samples t tests for D–H;