Human in vitro-induced regulatory T cells display Dlgh1dependent and PKC- restrained suppressive activity
Alexandra Zanin-Zhorov1,3*, Sudha Kumari1,4, Keli L. Hippen2, Sarah C. Merkel2, Margaret L. MacMillan2, Bruce R. Blazar2, and Michael L. Dustin1,5*
1
Molecular Pathogenesis Program, Skirball Institute of Biomolecular Medicine, Department of
Pathology, New York University School of Medicine, New York, NY10016, USA 2
University of Minnesota Cancer Center and Department of Pediatrics, Division of Blood and
Marrow Transplantation, Minneapolis, MN 55455, USA 3
Current address is Kadmon Corporation, LLC, New York, NY 10016, USA
4
Current address is Koch institute of Integrative Cancer Research, MIT, Cambridge, MA-02139,
USA 5
Current address is Kennedy Institute of Rheumatology, University of Oxford, Oxford OX3 7FY,
UK *
Corresponding authors: Alexandra Zanin-Zhorov PhD:
[email protected]
Michael L Dustin PhD:
[email protected]
Running title: Dlgh1 and PKC- oppositely controls in vitro-induced human Tregs
Supplementary Figure 1
Characteristics of expanded T cell subsets: a
Summary (n=5) 10000
80
Fold Expansion
%Foxp3+25+
100
60 40 20 0
nTreg
iTreg
1000
100
Teff
1
nTreg
b Representative FACS plot examples CD4-gated
tTreg
CD4-gated
iTreg
CD4-gated
Teff
10
iTreg
Teff
a
tTreg iTreg Teff
Supplementary Figure 2
Dlgh1
actin
b
c
tTreg iTreg Teff
tTreg iTreg Teff
PTEN actin
actin pp38
d
tTreg
iTreg
Teff
pAKT actin
Supplementary Figure 3 a
tTreg siRNA: control
iTreg siRNA: control Dlgh1
Dlgh1
Dlgh1
Dlgh1 actin 47
b
17
Treg treatment: control siRNA Dlgh1 siRNA
100
% inhibition of IL-17 secretion
actin
6
80
2
c
iTreg siRNA:
control
Dlgh1
pten
67% inhibition
60 actin
40
24
20
8
0
d IFN-g (ng/ml)
1.0 0.8
0.6 0.4 0.2 0.0 None
0
0.1
1
10
AEB071 (M)
e 100
p
