Table SI: P1PK and GLOB blood group phenotypes and antigens
Phenotypes
Antigens
P1
P1, P, Pk
P2
P, Pk
p
none
P1 k
P1, Pk
P2 k
Pk
1 Table SII: Expression of Pk/Gb3 on RBCs and allelic discrimination
Pk %
P1 %
Genotype
(Phenotype)
P1PK blood
group controls
p
0.6
0
1 1
P1
17.5
87.7
P /P (P1)
P2
0.5
0
P2/P2 (P2)
Patients
1
1.0
0
P2/P2 (P2)
17
13.5
12.6
P1/P2 (P1)
18
22.9
21.9
P1/P1 (P1)
19
1.5
3.2
P2/P2 (P2)
20
8.3
17,1
P1/P2 (P1)
22
18.5
53.0
P1/P1 (P1)
23
1.1
0
P2/P2 (P2)
k
P /Gb3/CD77 and P1 antigen levels on RBCs are presented as percentage of
the RBCs in three well-studied controls with known RBC phenotype p, P1 and
P2 as well as in 7 STEC-HUS patients. Allelic discrimination for the P1 or P2
phenotype: C/C=P1, C/T=P1 heterozygote and T/T=P2.
2 HUS patient
Healthy control
Unlabeled
sample
Anti-C3
Isotype control
Figure S1. Flow cytometry demonstrating C3 on RBCs
Primary histogram from one patient and one control showing unlabeled sample, anti-C3 labeled and the isotype
control
A
B
CD35
MFI APC
100
MFI FITC
3500
150
3000
2500
50
0
Unstimulated
2000
Stx2
E
Factor H
2500
1500
Unstimulated
Stx2
Unstimulated
Stx2
CD235a and Factor H
40
25
20
15
10
30
20
10
5
0
3000
2000
MV 103/mL
% RBC with bound Factor H
D
CD59
4000
3500
200
MFI FITC
C
CD55
Unstimulated
Stx2
0
Unstimulated
Stx2
Figure S2. The effect of Stx2 on expression of complement inhibitors on RBCs
Complement inhibitors on RBCs were measured in unstimulated and Stx2-stimulated samples (n=4). The results are presented
as mean fluorescent intensity (A-C), percent RBCs with bound factor H (D) or RBC-derived microvesicles with factor H (E) in
Stx2 stimulated samples, depicted as median (bar) and individual values. No statistical significance was found for all four
complement regulators.