Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Legends to supplementary figures Supplementary Figure 1 Apoptosis of neural progenitors after irradiation is dependent on p53 status. Apoptotic cells in the subgranular zone (SGZ) of the dentate gyrus (a−f) are rarely observed in non-irradiated mice (a, c, e). At 8 hours after 17 Gy, numerous apoptotic cells with characteristic nuclear condensation and fragmentation using DAPI nuclear staining are seen in the SGZ (b) of p53 wildtype (+/+) mice. Mice heterozygous () for p53 appear to have an intermediate response (d) after 17 Gy, and the response was not observed in mice knockout () of the p53 gene (f). The number of apoptotic cells at 8 hours after 17 is radiation dose and p53 status dependent (table). Supplementary Figure 2 Radiation-induced apoptosis of endothelial cells in mouse brain is regulated by smpd1 gene. A TUNEL positive cell (arrow, a) is associated with a CD31-labeled microvessel endothelial cell (arrow, b; merged, c) in the mouse dentate gyrus at 8 h after 17 Gy. The number of CD31 positive apoptotic cells in mouse dentate gyrus at 8 h after irradiation is radiation dose (P<0.001, two-way ANOVA) and smpd1 genotype (P<0.001) dependent (d). Error bars represent means ± S.E.M. Supplementary Figure 3 Cells dissociated from neurospheres cultured from mouse brain demonstrate phenotypic markers of neural progenitors. They show dual immunostaining for nestin (a) and Sox2 (b; DAPI, c; merged, d). These cells also show immunoreactivity for musashi-1 (e; DAPI, f; merged, g), phenotypic markers of neural progenitors. 1 Supplementary Figure 4 Neural progenitors cultured from smpd1+/+, smpd1−/−, p53+/+ and p53−/− mice demonstrate multipotential ability and are able to differentiate in vitro into oligodendrocytes, astrocytes and neurons as revealed by immunocytochemistry (red) for galactocerebroside (GC, a−d), GFAP (e−h) and MAP2 (i−l) respectively. DAPI nuclear counterstaining is shown in blue. Supplementary Figure 5 Neural progenitors undergo apoptosis in vitro after irradiation. At 24 hours after 5 Gy, musashi (a) and Sox 2 (d) stained cells demonstrate characteristic nuclear condensation and fragmentation of apoptosis as shown by DAPI nuclear countstaining (arrows, b and e; merged, c and f) Supplementary Figure 6 Neural progenitors cultured from eGFP mice demonstrate multipotential ability and can differentiate into oligodendrocytes (green, eGFP, a; red, GC, b), astrocytes (eGFP, c; GFAP, d) and neurons (eGFP, e, g; MAP2, f; -III tubulin, tub III, h). Nuclei are counterstained using DAPI. Supplementary Figure 7 Neural progenitors from eGFP mice undergo apoptosis in vitro after irradiation. Compared to non-irradiated cells (a−c), a single dose of 5 Gy induces many apoptotic cells (d−f) that show characteristic nuclear condensation upon DAPI (e), and are also positive for TUNEL (f). A single dose of 5 Gy induces a significant increase in the number of apoptotic cells in neural progenitors cultured from eGFP mice (g). Results represented average of a minimum of 3 experiments, and a minimum of 500 cells counted per experiment. Error bars represent means ± S.E.M., * P<0.01, t-test, compared to non-irradiated controls. 2 Supplementary Figure 8 Neural progenitors from eGFP mice transplanted stereotactically into the mouse hippocampus are able to differentiate into neurons. A transplanted eGFP cell (arrow, a) demonstrates the neural progenitor phenotypic marker nestin (b; DAPI, c; merged, d). Two eGFP cells in the dentate gyrus (arrows, e) show immunostaining for DCX (arrows, f), a marker for neuronal progenitors (DAPI, g; merged, h), and an eGFP cell (arrow, i) has differentiated into a neuron and demonstrates immunoreactivity for the neuronal marker NeuN (arrow, j; DAPI, k; merged, l). - 3