Journal: Aging and Disease
Article Title: Mild Therapeutic Hypothermia Protects the Brain from Ischemia/Reperfusion Injury through Upregulation of iASPP
doi: 10.14336/AD.2017.0703
Figure Lengend Snippet: Mild therapeutic hypothermia decreased expression of ASPP1 and ASPP2 after stroke. (A) Representative protein bands of ASPP1 and ASPP2 from the Western blot. Actin served as a loading control. Mice subjected to normothermic or mild hypothermic stroke were euthanized at 4, 12, 24 and 72 h later. (B) Densitometric quantification of ASPP1 and ASPP2 in Western blot, n = 4 per group. Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, vs sham group under normothermia; # p < 0.05 vs sham group under mild therapeutic hypothermia; ♦ p < 0.05, vs. corresponding group under normothermia, by two-way ANOVA and Sidak’s test. (C) Representative images showing expression of ASPP1 and ASPP2 (red) in neurons (green) of the cortex at 12 h after cerebral ischemia. DAPI was used as a nuclear marker.
Article Snippet: The following antibodies were used: rabbit anti-ASPP1 polyclonal antibody (1:200, Abbiotec, San Diego, CA, USA), rabbit anti-ASPP2 polyclonal antibody (1:200, Abbiotec), rabbit anti-iASPP polyclonal antibody (1:1000, Abcam, Hong Kong, China), rabbit anti-p53-upregulated mediator of apoptosis (Puma) (1:1000, Abcam), rabbit anti-Bax (1:200, Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), cleaved caspase-3 (1:1000, Cell Signaling Technology Inc. Beverly, MA, USA) and rabbit anti-Actin polyclonal antibody (1:2000, Santa Cruz Biotechnology Inc.).
Techniques: Expressing, Western Blot, Marker