Supplementary Figure Legends
Figure S1 Targeting vector for the inducible deletion of the DNase II gene.
The wild-type DNase II gene, targeting vector, targeted allele (3 lox), flox allele,
Cre-mediated deletion allele (DNase II), and previously established DNase
II-null allele are shown schematically. Black boxes represent exons of the
DNase II gene. Closed triangles represent the loxP sequence. The
PGK-promoter-driven neo gene and DNA coding for the diphtheria toxin A
fragment (DT-A) are indicated by boxes. The probe used for Southern
hybridization in Fig. S2 is indicated by a filled box below the wild-type allele.
Some recognition sites for major restriction enzymes (B, Bam HI; E, Eco RV;
Sm, Sma I; H, Hind III; X, Xho I; SI, Sac I; Nc, Nco I; Nr, Nru I; Xm, Xmn I, and
SII, Sac II) are indicated. The recognition sites destroyed during vector
construction or recombination are in parenthesis. A 1.0-kb size marker is shown.
Figure S2 Inducible deletion of the DNase II gene. Left panel, chimeric mice
generated with an ES clone carrying the DNase II3lox/+ allele were mated with
C57/BL6 mice. These mice were crossed with transgenic mice carrying E2a-Cre
to generate mice carrying the DNase IIflox/+ allele. Tail DNA was prepared from
DNase II+/+ (lane 1) and DNase II3lox/+ (lane 2) mice, digested with Sac I, and
subjected to Southern hybridization with the probe. Bands for the wild-type (WT)
and targeted allele (3lox) are indicated by arrowheads. Right panel, DNase
IIflox/-Mx1-CreT (lane 3) or DNase II+ /+Mx1-CreT mice (lane 2) at age 4-6 weeks
were treated with poly(I:C). Two months later, DNA was prepared from the
spleen, digested with Nco I, and analyzed by Southern hybridization. DNA from
untreated DNase IIflox/+ mice (lane 1) was analyzed as a control for the wild-type
and lox alleles. Bands for the conditionally deleted allele (delta), null allele (null),
and wild-type/ floxed allele (WT or flox) are indicated by arrowheads.
Figure S3 No expression of DNase II mRNA in poly(I:C)-treated mice. Four
to six-week-old DNase IIflox/- mice were treated with poly(I:C), and RNA was
prepared from the bone marrow and spleen at the indicated time. DNase II
mRNA was quantified by real-time PCR, and expressed as a value relative to
-actin. There were 3-5 mice in each group, and their average values are plotted
with S.D.(bars).