![Page 1: Immunoglobulin V regions diversify by gene conversion in DT40 cells](https://reader034.vdocuments.us/reader034/viewer/2022051622/5681552d550346895dc3074f/html5/thumbnails/1.jpg)
Immunoglobulin V regions diversify by gene conversion in DT40 cells
Able to assay of sIgM+status by flow cytometry
V V V V V J C1
V V V V V* J C1
V
V
V*
![Page 2: Immunoglobulin V regions diversify by gene conversion in DT40 cells](https://reader034.vdocuments.us/reader034/viewer/2022051622/5681552d550346895dc3074f/html5/thumbnails/2.jpg)
Chromatin features of pseudogene donors
Histone acetylation
H3.3 variant
![Page 3: Immunoglobulin V regions diversify by gene conversion in DT40 cells](https://reader034.vdocuments.us/reader034/viewer/2022051622/5681552d550346895dc3074f/html5/thumbnails/3.jpg)
10000
1000
100
10
0
0 400 600 800 1000200
10000
1000
100
10
0
0 400 600 800 1000200
Forward scatter Forward scatter
s IG
M
s IG
M
GFP-LacI HIRA-LacI
Effects on diversification
Med
ian
sIg
M
loss
1.0 1.7 11.0 8.4
0
2
4
6
8
10
12
14
GFP-LacI HIRA Cl18 HIRA-LacI VP16-LacI
![Page 4: Immunoglobulin V regions diversify by gene conversion in DT40 cells](https://reader034.vdocuments.us/reader034/viewer/2022051622/5681552d550346895dc3074f/html5/thumbnails/4.jpg)
Long tract GC77%
Short tract GC3%
Point Mutation20%
Insertion0%
Deletion0%
GFP-LacI
71
Long tract GC66%
Short tract GC21%
Point Mutation11%
Insertion2%
Deletion0%
HIRA-LacI
137
VP16-LacI
Long-tract GC45%
Short-tract GC29%
Point Mutation13%
Deletion0%
Insertion13%
31
LacI-HP1Long tract GC
3%Short tract GC
17%
Point Mutation58%
Insertion8%
Deletion14%
36
![Page 5: Immunoglobulin V regions diversify by gene conversion in DT40 cells](https://reader034.vdocuments.us/reader034/viewer/2022051622/5681552d550346895dc3074f/html5/thumbnails/5.jpg)
Long tract GC77%
Short tract GC3%
Point Mutation20%
Insertion0%
Deletion0%
GFP-LacI
71
Long tract GC66%
Short tract GC21%
Point Mutation11%
Insertion2%
Deletion0%
HIRA-LacI
137
VP16-LacI
Long-tract GC45%
Short-tract GC29%
Point Mutation13%
Deletion0%
Insertion13%
31
LacI-HP1Long tract GC
3%Short tract GC
17%
Point Mutation58%
Insertion8%
Deletion14%
36
![Page 6: Immunoglobulin V regions diversify by gene conversion in DT40 cells](https://reader034.vdocuments.us/reader034/viewer/2022051622/5681552d550346895dc3074f/html5/thumbnails/6.jpg)
GFP-LacI-11 HIRA-LacI-19
Effect of HIRA on chromatin
Cell 1
Cell 2
nucleosomedeposition
Cell 1
Cell 2
~150 bp
MNase
![Page 7: Immunoglobulin V regions diversify by gene conversion in DT40 cells](https://reader034.vdocuments.us/reader034/viewer/2022051622/5681552d550346895dc3074f/html5/thumbnails/7.jpg)
VP16-LacI-10GFP-LacI-11
AcH4 IgG AcH4 IgG
V17
Ova
Enrichment: 4.5 12.4
GFP-LacI VP16-LacI
Effect of VP16 on chromatin