Figure 8.

BDNF treatment increases SK2 serine phosphorylation. A, B, Western blots of (1) phosphoserine immunoprecipitates from homogenates of control (con) and BDNF-treated hippocampal slices (lanes 1, 2, 5, 6) and (2) COS7 cell lysates (lanes 3/4, 7/8) that had been probed with intact anti-SK2 (Alomone Labs) (A) or anti-SK2 that had been preabsorbed with antigen peptide (B). As seen in A, two sharp bands of SK2 immunoreactivity within the COS7 lysate (lanes 3/4; at 105 and 45–50 kDa) correspond with phosphoprotein bands in the hippocampal homogenates that become markedly more robust with BDNF treatment. B shows that antigen-preabsorbed antibody no longer detects (1) these bands in the COS7 lysate or (2) enhanced phosphoserine immunoreactivity at these molecular weights in samples of BDNF-treated hippocampal tissue. C, Western blot of SK2 immunoprecipitates from homogenates of control and BDNF-treated slices that were probed with anti-SK2 (Alomone Labs) show that BDNF does not influence levels of total SK2 immunoreactivity; the two lanes at left and right were loaded with 30 and 10 μl of sample, respectively. IP, Immunoprecipitates; WB, whole brain.