(A) The immune-precipitates with nonspecific IgG (N.S. IgG), or antibody against NMDA receptor NR2B subunit (anti-NR2B) in the membrane extracts (5 mg proteins) from the cortex of mice 2 hr after operation for sham or MCAO (Ischemia) were stained with Coomassie blue.
(B) Base peak chromatogram from trypsin-digested products of a selected protein band (arrow head in [A]).
(C and D) The membrane extracts (1 mg proteins) from the cortex of mice 2 hr after sham (lane 1) or MCAO (lane 2) were precipitated with nonspecific IgG (IgG) or anti-NR2B (C) or anti-DAPK1 (D) and probed with anti-DAPK1, anti-NR1, anti-PSD95 or anti-NR2B. Input: 20 μg of protein was loaded in each lane except the PSD95 lane, in which 10 μg proteins were loaded.
(E–G) DAPK1+/+ (E) and DAPK1−/− (G) cultured neurons were stained with anti-DAPK1 (red) and anti-NR2B (green). A high magnified image (F) from a selected area in (E). In panels A–G, similar results were observed in each of the four experiments.
See also Figure S1.