Figure 1. Effects of low-dose chronic treatment with I2-IR
LSL60101, Donepezil and co-administration on cognitive status in 5XFAD
and WT mice. (a) Scheme of experimental design. Results of
NORT: Discrimination index calculated by using exploration time for
novel and familiar object (b) in the short-term term memory
test session (2h) (WT, n=10-12 per group; 5XFAD n=11-12 per group)(c) in the long-term memory test session (24h) (WT, n=10-12 per
group; 5XFAD n=10-12 per group). Results of MWM : (d)Distance to target (platform) (cm) during the training session (WT
n=10-11 per group; 5XFAD n=10-12 per group) (e) Quadrant
preference in the test session as time (%) spent in each quadrant WT
n=9-12 per group; 5XFAD n=10-12 per group) (f) Time (%) spent
in platform quadrant in the test session (WT n=9-12 per group; 5XFAD n=
10-12 per group) (g) Distance to target (platform) (cm) in the
test session (WT n=9-12 per group; 5XFAD n=10-12 per group) (h)Representative images of trajectory during memory test. Bars represent
mean±SEM. one-way and two-way ANOVA with Tukey post hoc analysis,
*P<0.05.
Figure 2. Effects of low-dose chronic treatments I2-IR LSL60101,
Donepezil and co-administration on behavioral and social status in 5XFAD
mice and WT controls. Results of OF: (a) Locomotor activity measured as
distance travelled (cm) WT n=10-12 per group; 5XFAD n=11-12 per group(b) Time spent in the center (sec) (WT n=10-12 per group; 5XFAD
n=9-11 per group) (c) number of rearings (WT n=11-12 per group;
5XFAD n=10-12 per group). Results of EPM: (d) Time (%) spent
in Open Arms (WT n=11-12 per group; 5XFAD n=10-12 per group)(e) Time (%) spent in closed arms (WT n=11-12 per group; 5XFAD
n=10-12 per group) (f) number of rearings (WT n= 11-12 per
group; 5XFAD n=11-12 per group). Results of TCT: (g) Entries in
chambers (A1, A2) in the habituation phase (n) (WT n=11-12 per group;
5XFAD n=11-12 per group) (h) Time in chambers (Empty, Stranger)
during the test session (WT n=10-11 per group; 5XFAD n=9-11 per group)(i) time of interaction with intruder (sec) in the test session
(WT n=11-12 per group; 5XFAD n=10-11 per group). Bars represent
mean±SEM. Student’s t-test or two-way ANOVA with Tukey post hoc
analysis, *P<0.05;
Figure 3. Effects of low dose chronic treatments I2-IR LSL60101,
donepezil and co-administration on AD hallmarks in 5XFAD mice. (a)Histological images of amyloid plaques stained with Thioflavin-S and
quantification of (b) number of the plaques in cortex and
hippocampus (c ) and area (%) covered by plaques in the DG area
of the hippocampus in the 5XFAD mice. Representative Western blot and
quantifications (d-f) for β-Amyloid p-Tau(Ser404) and
p-Tau(ser396) in the hippocampus of 5XFAD mice. Values in bar graphs are
adjusted to 100% for protein levels of the control WT control or the
control 5XFAD. Bars represent mean±SEM. Student’s t-test or one-way
ANOVA with Tukey post hoc analysis, *P<0.05; n=5-6 per group.
Figure 4. Effects of low dose chronic treatments I2-IR LSL60101,
donepezil and co-administration on Aβ pathology and APP processing in
5XFAD mice. Representative Western Blots and Quantifications for(a-g) CTFs, p-APP, sAPPβ, sAPPα, BACE 1, ADAM10 in the
hippocampus of 5XFAD mice. Values in bar graphs are adjusted to 100%
for protein levels of the control WT control or the control 5XFAD.
Representative gene expression in the hippocampus of the 5XFAD mice for(h) APP and (i) IDE, Nep. Gene
expression levels were determined by real-time PCR. Bars represent
mean±SEM. Student’s t-test or one-way ANOVA with Tukey post hoc
analysis, *P<0.05; n=5-6 per group.
Figure 5. Effects of low dose chronic treatments I2-IR LSL60101,
donepezil and co-administration on microgliosis and inflammatory markers
in 5XFAD mice. (a) Representative images of GFAP immunostaining and
quantification (b-d) in DG, CA1, CA3 areas of the hippocampus
of the 5XFAd mice. Representative gene expression in the hippocampus of
the 5XFAD mice for (e) Il-1β, Il-6, Ccl12, (f)for Trem2 . Gene expression levels were determined by real-time
PCR. Bars represent mean±SEM. Student’s t-test or one-way ANOVA with
Tukey post hoc analysis, *P<0.05; n=5-6 per group.
Figure 6. Effects of low dose chronic treatments I2-IR LSL60101,
donepezil and co-administration on astrogliosis and synaptic plasticity
in 5XFAD mice. (a) Representative images of GFAP immunostaining and
quantification (b-d) in DG, CA1, CA3 areas of the hippocampus
of the 5XFAd mice. Representative western blot and quantifications(f-h) for PSD95 and SYN in the hippocampus of 5XFAD mice.
Values in bar graphs are adjusted to 100% for protein levels of the
control WT control or the control 5XFAD. Bars represent mean±SEM.
Student’s t-test or one-way ANOVA with Tukey post hoc analysis,
*P<0.05; n=5-6 per group.