Figure S6 from A Metabolic Gene Survey Pinpoints Fucosylation as a Key Pathway Underlying the Suppressive Function of Regulatory T Cells in Cancer

Abstract

<p>Additional evidence of the effect of systemic administration of a fucosylation inhibitor on reducing tumor immunosuppression. Related to Figure 6. CT26 tumor-bearing mice, were treated with 2FF or saline treatment. (A) Representative histogram of PD-1 expression in Tregs derived from CT26 tumors treated with 2FF or saline. (B) Percentage of Tregs out of CD45+ in the spleens of CT26 tumor-bearing mice upon 2FF or saline treatment. (C) Percentage of CD4+Foxp3- cells out of CD45+ in the spleens of CT26 tumor-bearing mice upon 2FF or saline treatment. (D) Percentage of CD8+ out of CD45+ in the spleens of CT26 tumor-bearing mice upon 2FF or saline treatment. (E) PD-1 MFI in Tregs in the spleens of CT26 tumor-bearing mice upon 2FF or saline treatment. (F) Percentage of PD-1+ out of Tregs cells in the spleens of CT26 tumor-bearing mice upon 2FF or saline treatment. (G) PD-1 MFI in CD4+Foxp3- T cells in the spleens of CT26 tumor-bearing mice upon 2FF or saline treatment. (H) Percentage of PD-1+ out of CD4+Foxp3- T cells in the spleens of CT26 tumor-bearing mice upon 2FF or saline treatment. ¬(I) PD-1 MFI in CD8+ T cells in the spleens of CT26 tumor-bearing mice upon 2FF or saline treatment. (J) Percentage of PD-1+ out of CD8+ T cells in the spleens of CT26 tumor-bearing mice upon 2FF or saline treatment. (K) MFI AAL in macrophages (F4/80+), neutrophils (Ly6G+) and cancer cells in the CT26 tumors. (L) Percentage of neutrophils Ly6G+ (out of CD45+) in the CT26 tumors. (M) Percentage of CT26 CD90.1+ cancer cells out of alive in the CT26 tumors. (N) Percentage of Tregs (out of CD45+) in the CT26 tumors. (O) Percentage of CD4+Foxp3- T cells (out of CD45+) in the CT26 tumors. (P) Percentage of macrophages F4/80+ (out of CD45+) in the CT26 tumors. (Q) Percentage of MHCIIHIGH (out of F4/80+ macrophages) in the CT26 tumors. (R) Percentage of alive, MHCIIHIGH, CD80, CD86, PD-L1 and CD206 (out of F4/80+ macrophages) upon ex vivo treatment of bone marrow-derived macrophages (M0) with DMSO or 2FF. (S) Percentage of CD8+ T cells (out of CD45+) in the CT26 tumors. (T) Percentage of Tregs out of CD4+ T cells in the blood of healthy mice 3 days upon treatment with anti-CD25 or anti-IgG control. (U) Percentage of Tregs out of CD4+ T cells in CT26 tumors at end-stage. (V) Relative percentage of F4/80+ cells out of CD45+ cells versus the anti-CD25 + saline group in the tumor of CT26 tumor-bearing mice. (W) Relative percentage of MHCIIHIGH out of F4/80+ cells versus the anti-CD25 + saline group in the tumor of CT26 tumor-bearing mice. For V - W, given the smaller tumor size upon anti-CD25 treatment, the effect of 2FF administration in Treg-depleted mice is shown in comparison to its own saline control. (X) Flow chart of the PD-1 reconstituted Tregs 3 days upon transduction, where GFP is the transduction marker. Statistical analysis was performed using unpaired two-tailed t-test (Fig. S6B-J, L-Q, S-T and V-W) or 2-way ANOVA with Sidak’s multiple comparisons test (Fig. S6K, R and U) and are represented as mean ± SEM (ns, p > 0.05, * p  0.05, ** p  0.01 and *** p  0.001).</p>