1. 0 Gy NSC NSC 5 Gy NSC NSC NSC NSC NSC NSC NSC NSC NSC NSC NSC NSC
Media
transfer
Detection of cell
cycle and apoptosis
Differentiation
(without FGF2
and EGF)
0 Gy
-24 h
0 h
24 h
48 h
72 h
10 days
NSC
NSC
NSC
200x
Control-2
NSC
NSC
5 Gy
(Control-1)
Apoptosis
Control-2 6%
NSC
NSC
NSC
(Irradiated)
Relative
Survival 100%
Recipients for
media transfer
Bystander
(+ 24/48 h)
NSC
Apoptosis
NSC
NSC
(Control-2)
18%
Bystander
Relative
Survival 84%
NSC
NSC
NSC
(Bystander)
Directly irradiated (5 Gy)
Suppl. Fig. 2. Radiation-induced bystander effects in neural stem cells (NSC): design of experiments with non-treated and irradiated media transfer. NSC were non-treated (control-1) or irradiated at 5 Gy. 24 h after irradiation, control-1 (non-irradiated) media and irradiated media were transferred to naive non-targeted NSC (control-2) and bystander cells, respectively, which were cultured an additional 24 or 48 h. Then cell cycle- apoptosis analysis was performed using flow cytometry. Additional experiments were performed for induction of neuronal differentiation of control, bystander and directly irradiated cells using the differentiation medium. Nestin, early neuroprogenitor marker, (red); Doublecortin, neuronal marker (green) have been used for evaluation of neuronal differentiation.
Relative
Survival 11%