Maarten van der Doelen

Chapter 3

A

Checkpoint panel 1/2

Fluorescence minus one Sample

CD8

FSC-A

SSC-A

Vialbility dye

Vialbility dye

FSC-A

CD3

CD4

FSC-H

CD3

CD4

CD4

CD4

CD4

CD4

CD4

PD-1

CTLA-4

ICOS

PD-L1

TIM-1

TIM-3

B

T cell panel

Effector/memory subsets

CD8

CCR7 CCR7

FSC-A

SSC-A

Vialbility dye

CD4

CD45RO

FSC-H

CD3

FSC-A

Regulatory T cells

CD4

CD4

FSC-W

FoxP3

CD25

CTLA-4

CD45RO

C

M-MDSCs

MDSC panel

CD15

FSC-W

HLA-DR

CD14

CD11b

PD-L1

eMDSCs

FSC-A

SSC-A

Vialbility dye

FSC-H

CD14

FSC-A

CD14

FSC-W

HLA-DR

Lin

CD33

PD-L1

Supplementary figure 1. Gating strategy. A . Checkpoint panel 1 and 2. Gates for checkpoint expression on CD4 + (upper gate) and CD8 + cells (lower gate) are shown in the same graph. B . T cell panel. C . MDSC panel. Fluorescence minus one (FMO) controls were used to gate the population that expressed checkpoint molecules and HLA-DR.

78