Supplementary Files
Title: Therapeutic Intervention of Learning and Memory Decays by Salidroside
Stimulation of Neurogenesis in Aging
Running Title: Treatment of Learning and Memory Decays in Aging
Authors: Huijuan Jin1,5*, Lei Pei1,5*, Xiaogang Shu2,5*, Xin Yang1,5, Tianhua Yan3,
Yan Wu2,5, Na Wei1,5, Honglin Yan1,5, Shan Wang1,5, Chengye Yao1,5, Dan Liu4,5,
Qing Tian5, Lin Wang2,5#, and Youming Lu1,5#
1
Department of Physiology, Tongji Medical College, Huazhong University of Science
and Technology, Wuhan, 430030, China
2
Translational Medical Research Center, the Union Hospital, Huazhong University of
Science and Technology, Tongji Medical College, Wuhan, 430022, China
3
Department of Physiology, China Pharmaceutical University, No. 24, Tong Jia Xiang
Street, Nanjing 210009, China
4
Department of Genetics, Tongji Medical College, Huazhong University of Science
and Technology, Wuhan, 430030, China
5
Institute of Brain Research, Huazhong University of Science and Technology,
Wuhan, 430030, China
Supplementary Figure 1
a
b
Differentiation
Proliferation
Proliferation
Vehicle
Salidroside
50
Vehicle
Salidroside
50
Cells/0.5cm2
Cells/0.5cm2
Differentiation
25
25
*
0
24h
72h
4d
8d
0
24h
72h
4d
8d
Effects of Salidroside at different concentrations on neural progenitor cell survival.
a, Salidroside at 0.5 μM does not alter differentiation of rat fetal neural stem cells (NSCs)
into neurons. Bar graph shows proliferation (24 and 72 h) and differentiation (4 and 8 days)
of NSCs in the presence of vehicle or 0.5 µM Salidroside. The number of proliferating
cells (nestin+/ki67+ cells) and differentiated cells (MAP2+/DCX+ or NeuN/TUJ1+ cells)
from vehicle and Salidroside groups are summarized (n = 4). Above the graph indicates 72
h after proliferation (arrow), the culture medium was replaced with neuronal basal medium.
b, Salidroside at 5 μM concentration enhances differentiation of NSCs into neurons. Bar
graph shows show proliferation (24 and 72 h) and differentiation (4 and 8 days) of NSCs in
the presence of vehicle or 5 µM Salidroside. The number of proliferating cells
(nestin+/ki67+ cells) and differentiated cells (MAP2+/DCX+ or NeuN+/TUJ1+ cells) from
vehicle and Salidroside groups are summarized (n = 4, *p<0.01).
Time
3000 (days)
2000
1000
Supplementary Figure 2
Salidroside
BrdU S1 S2
S3
S4
5d
Vehicle
Salidroside
12 h
200 µm
200 µm
72 h
200 µm
200 µm
7d
200 µm
200 µm
14 d
200 µm
200 µm
Vehicle
Brdu+ cells
3000
Salidroside
2000
1000
0
0
7
Time (days)
14
Photomicrographs of the BrdU+ nuclei in adult mice treated with vehicle or
Salidroside. An illustration above the images indicates the experimental
schedule 12 h (S1), 24 h (S2), 7 days (S3), or 14 days (S4) after BrdU
injection .
Supplementary Figure 3
a
Salidroside BrdU S1 S2
S3
S4
5d
Vehicle
Salidroside
2000
1000
Vehicle
Salidroside
3000
Brdu+ cells
Brdu+ cells
3000
b
2000
*
1000
0
0
0
7
Time (days)
0
14
7
Time (days)
14
d
c
Vehicle
Salidroside
2000
1000
Vehicle
Salidroside
3000
Brdu+ cells
Brdu+ cells
3000
2000
1000
0
0
0
7
Time (days)
14
0
7
Time (days)
14
Effects of Salidroside on the survival of endogenous neural progenitor cells (NPCs) in the dentate
gyrus during aging. a-c, The effects of Salidroside at a dose of 1 mg/kg (a), 10 mg/kg (b) (per dose
per day for 5 consecutive days) or 5 mg/kg (c) (per dose per day for 2 consecutive days) on the fate
of endogenous progenitor cells in the dentate gyrus of aging mice was examined. The number of
BrdU+ cells was assessed and summarized in the graphs. An illustration above the graph indicates the
experimental schedule 12 h (S1), 24 h (S2), 7 days (S3), or 14 days (S4) after BrdU injection (n = 11
mice per group, *p<0.01). d, The stimulatory effects of Salidroside on NPCs survival were not found
when BrdU was injected 7 days after the administration of Salidroside. In this study, multiple doses
(5 mg/kg per dose per day for 5 consecutive days) of Salidroside were administered. 7 days after the
administration, BrdU was injected. 12 h, 24 h, 7 days or 14 days after the BrdU injection, BrdU
labeled nuclei was examined, and summarized in a bar graph (n = 11 mice per group).