Marilen Benner

THREE TYPES OF TREG CONTROL PLACENTAL INFLAMMATION 75 3 SUPPLEMENTAL INFORMATION FSC-A SSC-A FSC-H FSC-W SSC-H SSC-W CD45 CD14 CD56 CD4 R1 (55%) R1 R2 (55%) R2 R3 (54%) R3 R4 (47%) R4 R5 CD4+ (5%) R5 (11%) CD8+ (4%) A B CD25 TIGIT CD4+ R6 TIGIT + CD25 HI PD1 HI R6 Teff PD1 CD4 CD45 CD8 CD45RA CD127 FOXP3 C Supplementary Figure S1. FACS gating strategy. Related to Figure 1. A) CD4+ and CD8+ T cells were identified by selecting live cells (R1), duplicates were excluded (R2, R3), and CD45+CD56−CD3+CD4+ (CD4+) and CD45+CD56−CD3+CD8+ (CD8+) T cells were selected. B) Based on CD25, PD1 and TIGIT expression, CD4+ T cells were further separated into CD25 HI , PD1 HI , TIGIT+ and Teff cells. Representative FACS plots of C) FOXP3, CD45RA and CD127 expression and D) PD1 and CD25 expression in CD25 HI , PD1 HI and TIGIT+ cells in a 6-12wk decidual sample. E) Graph depicts the expression of FOXP3 (MFI) within CD25 HI and TIGIT+ cells in blood, decidua 6-12wk, decidua basalis >37wk and d.parietalis >37wk. n=8-10. Bars represent median and interquartile range; n= 8-10; *P<0.05, **P<0.01, ***P<0.001. E TIGIT PD1 CD25 D PD1 CD25 ● CD25 HI ● PD1 HI ● TIGIT+ MFI FOXP3 0 5 0 0 1 0 0 0 1 5 0 0 2 0 0 0 2 5 0 0 * *** ** ** TIGIT+ CD25 HI Supplementary Figure S1. FACS gating strategy. Related to Figure 1. (A) CD4+ and CD8+ T cells were identified by selecting live cells (R1), duplicates were excluded (R2, R3), and CD45+CD56−CD3+CD4+ (CD4+) and CD45+CD56−CD3+CD8+ (CD8+) T cells were selected. (B) Based on CD25, PD1 and TIGIT expression, CD4+ T cells were further separated into CD25 HI , PD1 HI , TIGIT+ and Teff cells. Representative FACS plots of (C) FOXP3, CD45RA and CD127 expression and (D) PD1 and CD25 expression in CD25 HI , PD1 HI and TIGIT+ cells in a 6-12wk dec dual sample. (E) Graph depicts the expression of FOXP3 (MFI) within CD25 HI nd TIGIT+ cells in blood, decidua 6-12wk, decidua bas lis >37wk a d d.parietalis >37wk. n=8-10. Bars represent median and interquartile range; n= 8-10; *P<0.05, **P<0.01, ***P<0.001.