DR-region of Na+/K+ ATPase is a target to treat excitotoxicity and stroke

a, b Western blots showing that protein levels of cortical tissue lysate (a) and plasma membrane (b) of total NKAα (a) and NKAα1 were markedly reduced in NKAα1+/？ mice, but not NKAα2 and NKAα3. n？=？4–5, Unpaired t test, two-tailed, *p<？0.05, **p？<？0.01 vs NKAα1+/+ group. c Infarct volumes induced by tMCAO (60？min ischemia followed by reperfusion). TTC stained sections of a representative brain from each group. n？=？12 for NKAα1+/+ group, n？=？10 for NKAα1+/？ group, Unpaired t test, two-tailed, *p？<？0.05 vs NKAα1+/+ group. d–g Generation of DR-Ab. d Schematic illustration showing the extracellular DR region of NKAα1subunit that binds DR-Ab. e ELISA of DR-Ab. f, g Representative western blots showing NKAα1 detected from membrane lysate in mouse brain (f), HEK293 and SH-SY5Y cells (g) using DR-Ab. h Infarct volumes induced by tMCAO (60？min ischemia followed by reperfusion). TTC stained sections of a representative brain from each group. DR, DR-Ab (30？μg/mouse, i.c.v.) pretreated group; NS, IgG purified from normal rat serum. n？=？10 for NS group, n？=？7 for DR group, Unpaired t test, two-tailed, *p？<？0.05 vs NS group. i Left panel, illustration of penumbra area in tMCAO model; middle panel, NKA activity in sham and sham+NS group, n？=？4/group, t-test; right panel, NKA activity in the penumbra area of the tMCAO mice. n？=？5/group. One-way ANOVA followed by Bonferroni’s test, *p？<？0.05, **p？<？0.01. j Infarct volumes induced by tMCAO (60？min ischemia followed by reperfusion). TTC stained sections of a representative brain from each group. DR, DR-Ab (30？μg/mouse, i.c.v.) post-treated group; NS, IgG purified from normal rat serum. n？=？9 for NS group, n？=？11 for DR group, Unpaired t test, two-tailed, *p？<？0.05 vs NS grou