The puzzling CDW -phase of 1 T-TiSe2 from a TR-ARPES point ...
Puzzling Low Surface Expression of H-DreI-B7 in IgL-targeted Clones Dre #4Dre #3Dre #2 Dre #3 Dre #2...
5
Puzzling Low Surface Expression of H-DreI-B7 in IgL-targeted Clones 10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4 5.69 0.37 1.02 92.9 Dre #4 10 0 10 1 10 2 10 3 10 4 0 20 40 60 80 100 10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4 0.12 11.7 86.6 1.57 10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4 1.41 0.12 0.63 97.8 Dre #3 Dre #2 10 0 10 1 10 2 10 3 10 4 0 20 40 60 80 100 10 0 10 1 10 2 10 3 10 4 0 20 40 60 80 100 Dre #3 Dre #2 IgM-FITC HA-APC dsOligo-Biotin/SAv-PE Cell count (%) unstained dsDre4 dsAni1 Dre #4 DT40 Ψ-/- AID R DT40 TetR WT
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Transcript of Puzzling Low Surface Expression of H-DreI-B7 in IgL-targeted Clones Dre #4Dre #3Dre #2 Dre #3 Dre #2...
Puzzling Low Surface Expression of H-DreI-B7in IgL-targeted Clones
100 101 102 103 104100
101
102
103
1045.69 0.37
1.0292.9
Dre #4
100 101 102 103 1040
20
40
60
80
100
100 101 102 103 104100
101
102
103
1040.12 11.7
86.61.57
100 101 102 103 104100
101
102
103
1041.41 0.12
0.6397.8
Dre #3Dre #2
100 101 102 103 1040
20
40
60
80
100
100 101 102 103 1040
20
40
60
80
100
Dre #3Dre #2IgM-FITC
HA
-APC
dsOligo-Biotin/SAv-PE
Cell
cou
nt
(%)
unstained dsDre4dsAni1
Dre #4
DT40 Ψ-/- AIDRDT40 TetR WT
Hypermutation optimized-DreI-B7/Hypermut2 clones
0 102 103 104 105
0
102
103
104
105 0.18 0.022
0.3299.5
0 102 103 104 105
0
102
103
104
105 6.37 70.8
20.32.49
0 102 103 104 105
0
102
103
104
105 2.72 82.7
140.6
0 102 103 104 105
0
102
103
104
105 5.57 89.1
4.330.97
0 102 103 104 105
0
102
103
104
105 0.087 10.3
85.73.99
0 102 103 104 105
0
102
103
104
105 0.83 73
25.50.69
0 102 103 104 105
0
102
103
104
105 0.5 36.4
61.71.42
0 102 103 104 105
0
102
103
104
105 0.52 85.7
13.80.071
3D5 3D5
3F4 HDreI 5A8 HDreI 3F5 HDreI 5F8 HDreI
5B8 HDreI 5B12 HDreI
3 weeks in culture:
1 week in culture:I-AniI/Hypermut2 control
Better surface expression, but no integration into rearranged IgL allele
IgM-FITC
HA
-tag
-AP
C
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
DT40 H-DreI C4
HyperDreI 5A8 HyperDreI 5F8 HyperDreI 4E12 HyperDreI 5E6
dsDre4-BT:SAv-PE
Cel
l cou
nt (
% o
f max
imum
)
Good recognition of H-DreI target sequence
Control cells
dsDre4 staining corresponds well to HA-tag staining
unstained dsDre4
Flow cytometry analysis of CTLMA2 target
5’ TGAGGAGGTTTCTCTGTAA 3’5’ AGAGGACCTTCATCTGTCA 3’
I-AniI WT:I-AniI CTLMA2:
14.0
6.3
7.8
1.0
1.0
1.0
1.0
1.0
1.0
2.6
2.4
1.3
MFl B10MFl DT40
MFl DT40 = 1.0
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
0 102 103 104 1050
20
40
60
80
100
DT
40B
3 I-
Ani
IB
10 I-
Ani
I3D
5 I-
Ani
IdsAni1 wt dsAni1 +1C+2A
dsOligo-Alexa647
Ce
ll co
un
t (%
of m
axi
mu
m)
Flow cytometry cleavage assay for CTLMA2 target
FACS cleavage assay
0.50
1.00
1.50
2.00
2.50
DT40 B3 B10 3D5
Ale
xa64
7/P
E f
luo
resc
ence
rat
io
dsAni1 wt
dsAni1 +1C+2A
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