Figure 3
In vitro kinase activity of Fps and Fer isolated from neonatal mouse brains. Fps and Fer were immunoprecipitated from neonatal mouse brain lysates from the indicated strains (wt, fps-/-, ferDR/DRor fpsKR/KR/ferDR/DR) with anti-Fps/Fer antibody. The immune precipitates were processed for immune complex kinase assays with or without added ATP as described in Materials and Methods and tubulin was added as an exogenous substrate. (A), Beads containing immunoprecipitated Fps and Fer were immunoblotted with anti-pY to reveal the degree of Fps/Fer in vitro autophosphorylation (upper). The blot was stripped and reprobed with anti-Fps/Fer antibody (lower). (B), Supernatants containing tubulin were immunoblotted with anti-pY to reveal the degree of tubulin phosphorylation (upper). The blot was then stripped and reprobed with anti-tubulin antibody (lower).