Supplementary information
Statistics
Statistical evaluation of the quantitative RT-PCR data for the three Aph-1 family
members was performed by means of a univariate analysis of variance (ANOVA) with as
dependent variable the normalized transcript levels and as fixed factors the genotypes
(I/I, II/II and III/III) and the developmental time points (either ED or PND). For every
time point, data were further analyzed for significant differences between the three
genotypes using a one-way ANOVA and where appropriate a post hoc Bonferroni test.
Per genotype, the statistical analysis for differences in transcript levels during
development was performed by means of an independent samples T-test to compare the
two embryonic time points, or using a one-way ANOVA and a post hoc Bonferroni test
for the postnatal time points. A probability of P < 0.05 was considered statistically
significant. For the longitudinal Western blot analyses, the data sets were statistically
analyzed by means of a univariate ANOVA with as dependent variable the APPCTF/APP-FL levels and as fixed factors the genotypes and the developmental time
points. Subsequent analysis using an independent samples T-test at every time point
revealed significant differences between I/I and III/III rat tissues. Per rat line, a one-way
ANOVA and where appropriate a post hoc Bonferroni analysis was used to identify
differences between developmental time points. All statistical analyses were performed
with the SPSS 12.0.1 software program (SPSS Inc., Chicago, Illinois, USA).
Supplemental table 1 Statistical support for Figure 1 by means of a univariate ANOVA
on longitudinal mRNA expression levels of Aph-1 family members in the hippocampus
of I/I, II/II and III/III rats (significant if P < 0.05; NS: no significant differences).
ED
PND
Aph-1b
Aph-1aS
Aph-1aL
geno: F(2,19) = 57.6
geno: NS
geno: NS
ED: F(1,19) = 9.8
ED: F(1,21) = 12.6
ED: F(1,22) = 18.9
geno*ED: F(2,19) = 4.5
geno*ED: NS
geno*ED: NS
geno: F(2,81) = 242.3
geno: NS
geno: NS
PND: F(1,81) = 26.8
PND: F(1,86) = 64.1
PND: F(1,86) = 20.0
geno*PND F(1,81) = 3.2
geno*PND: NS
geno*PND: NS
Supplemental table 2 Since the univariate ANOVA for Figure 1 yielded statistical
significance, we performed for the I/I, II/II and III/III rats over the embryonic time points
an independent samples T-test and over the postnatal time points a one-way ANOVA and
where appropriate a post hoc Bonferroni analysis. Significant differences between time
frames are indicated (significant if P < 0.05; NS: no significant differences).
ED
PND
independent
one-way ANOVA
Bonferroni
samples T-test
PND(0-4) < PND(22-100)
I/I
NS
PND: F(7,27) = 17.7
PND(9-12) < PND(35-60)
II/II
NS
PND: F(7,25) = 5.5
PND(0) < PND(22)
Aph-1b
PND(0-4) < PND(22-100)
III/III
ED13 < ED17
PND: F(7,26) = 17.2
PND(0-4) < PND(9)
PND(12) < PND(22-35)
Aph-1aS
Aph-1aL
I/I
NS
PND: F(7,27) = 19.1
PND0-60) < PND(100)
II/II
NS
PND: F(7,27) = 17.9
PND(0-60) < PND(100)
III/III
NS
PND: F(7,29) = 29.8
PND(0-60) < PND(100)
I/I
NS
PND: F(7,27) = 11.6
PND(0-9) < PND(35-60)
II/II
NS
PND: F(7,27) = 3.2
NS
III/III
NS
PND: F(7,29) = 11.4
PND(0-12) < PND(35-100)
Supplemental Table 3 Since the univariate ANOVA for Figure 1 yielded statistical
significance, we performed for every developmental time point a comparison of the
transcript levels of the I/I, II/II and III/III rats by means of a one-way ANOVA and where
appropriate a post hoc Bonferroni analysis. Significant differences between genotypes are
indicated (significant if P < 0.05; NS: no significant differences).
13
ED
17
0
4
9
12
PND
22
35
60
100
one-way ANOVA
Bonferroni
one-way ANOVA
Bonferroni
one-way ANOVA
Bonferroni
one-way ANOVA
Bonferroni
one-way ANOVA
Bonferroni
one-way ANOVA
Bonferroni
one-way ANOVA
Bonferroni
one-way ANOVA
Bonferroni
one-way ANOVA
Bonferroni
one-way ANOVA
Bonferroni
Aph-1b
Aph-1aS
Aph-1aL
geno: F(2,8) = 34.1
(I/I or II/II) < III/III
geno: F(2,9) = 34.0
(I/I or II/II) < III/III
geno: F(2,8) = 129.5
(I/I or II/II) < III/III
geno: F(2,8) = 52.0
(I/I or II/II) < III/III
geno: F(2,8) = 148.2
(I/I or II/II) < III/III
geno: F(2,8) = 90.9
(I/I or II/II) < III/III
geno: F(2,10) = 10.7
(I/I or II/II) < III/III
geno: F(2,8) = 135.5
I/I < II/II < III/III
geno: F(2,15) = 51.9
I/I < II/II < III/III
geno: F(2,8) = 295.3
I/I < II/II < III/III
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
-
NS
NS
NS
NS
NS
NS
NS
NS
NS
geno: F(2,8) = 5.2
NS
Supplemental table 4 Statistical support for Figure 2 by means of a univariate ANOVA
on longitudinal APP-CTF levels in the spinal cord and lung of I/I and III/III rats. Since
the univariate ANOVA yielded statistical significance, we performed an independent
samples T-test over the APP-CTF levels for every developmental time point, and for both
I/I and III/III rats a one-way ANOVA and post hoc a Bonferroni analysis. Significant
differences between genotypes or time frames are indicated (significant if P < 0.05; NS:
no significant differences).
spinal cord
univariate ANOVA
geno: NS
PND: F(4,30) = 15.6
geno*PND: NS
geno
I/I
III/III
PND
2
9
13
35
60
independent samples T-test
NS
NS
I/I > III/III
I/I > III/III
NS
one-way ANOVA
F(4,14) = 4.4
F(4,14) = 46.2
Bonferroni
PND(2) < PND(35)
PND(2-13) < PND35-60)
lung
univariate ANOVA
geno: F(1,30) = 13.1
PND: F(4,30) = 22.7
geno*PND: F(4,30) = 3.4
geno
I/I
III/III
PND
2
9
13
35
60
independent samples T-test
NS
NS
NS
I/I > III/III
I/I > III/III
one-way ANOVA
F(4,14) = 21.3
F(4,14) = 4.4
Bonferroni
PND(2-9) < PND(13-60)
NS