Cell - 8 September 2016

Supplemental Figures

100

80

60

40

20

0

Follicular

Frequency (%)

Marginal zone

Frequency (%)

WTGL

MutHL^121

HGLL^121

Splenocytes

120

90

60

30

Total numbers (x10 0

6 ) (^80) B cells
60
40
20
Total numbers (x10 0
6 )
Long homology arm Short homology arm
D4-1 J1-4
loxPACN loxPProm.VDJ
WT IGH locus
Targeting vector
Targeted IGH locus
Long homology arm Short homology arm
WT IGK locus J1-5
Targeting vector
Targeted IGK locus
loxPACN loxPProm.VJ
A
Prom.VJ
Prom.VDJ
VH and DH genes Mouse IGH constant genes
VH and DH genes Mouse IGH constant genes
VK genes Mouse IGK constant genes
VK genes Mouse IGK constant genes
IGHV 4-5901
93
IGLV 3-2102
93 94 93
IGHV 4-5901 IGLV 3-2102
GLHL121 MutHGLL 121
D
BC
CD23
CD21
IgM
IgD
Lambda
IgM
WT GLHL 121 MutHGLL 121
16.9
72.2
28.4
59.9
49
26.6
3.77 1.3 14.1
75.2
10.4
65.8
21.1
69.1
11.2
150
5
10
15
20
25
30
0
GLHL121 MutHGLL 121
Heavy chains Light chains
MZ MZ MZ
FO FO FO
Figure S1. Generation and Characterization of GLHL121and MutHGLL121 Knockin Mice, Related toFigure 1.
(A) Schematic representation of the targeting strategy used to generateIgVDJ(top) orIgVJ(bottom) knockin mice. The wild-type locus, the targeting vector and
the targeted locus are illustrated. The targeting vector contains long and short arms of homology to the wild-type mouse IGH or IGK loci indicated in gray; a self-
excising tACE-Cre/ Neo (ACN) cassette in green flanked by 2 loxP sites indicated in yellow. The corresponding promoters for the IGH or IGK are shown in blue,
and the humanVDJandVJare indicated in red and orange respectively. The excised genes in the wild-type locus are represented in black.
(B) Representative FACS plots showing frequencies of marginal zone (MZ) (CD21hi, CD23lo) and follicular (FO) (CD21lo, CD23hi) B cells (top); frequencies of IgM+
and IgD+B cells (middle); and frequencies of IgM+Lambda+B cells (bottom) in live, Dump
, B220+cells obtained from spleen homogenates of wild-type, GLHL 121
and MutHGLL121 knockin mice.
(C) Graphs show total numbers of splenocytes (upper left), total numbers of B cells (Live, Dump
, B220+) (upper right), frequencies of marginal zone (lower left)
and follicular (lower right) B cells in the spleens of wild-type, GLHL121 and MutHGLL121 knockin mice.
(D) Pie charts show heavy and light chain sequences amplified from single IgM+B cells isolated from naive GLHL121 and MutHGLL121 knockin mice. The number
in the center of the pie chart indicates the number of sequences amplified. Areas in colors represent the number of sequences corresponding to the knockin
humanV(D)Jgenes.