Fig. 2.

Expression of Fas and Fas ligand (A) or Fas–Fas ligand interaction (B) after CCI in mice over time [in hours (h)]. Contused or normal (uninjured control;C) cortex was subjected to Western blot analysis using polyclonal anti-Fas (M-20) or anti-Fas ligand (N-20). Fas and Fas ligand were constitutively expressed in uninjured cortex. Expression of Fas receptor was increased after CCI, whereas there was no change in Fas ligand expression. Bands corresponding to β-actin were of equal intensity in all lanes, suggesting equal protein loading. B, Fas–FasL interaction is increased after CCI. Fas ligand or Fas receptor was immunoprecipitated from cortical supernatant homogenates with the corresponding polyclonal antibodies and subjected to SDS-PAGE. Western blots were probed with monoclonal anti-Fas or anti-Fas ligand, respectively. A time-dependent increase in Fas–Fas ligand interaction was found. Robust interaction between Fas and Fas ligand was observed regardless of whether anti-Fas or anti-Fas ligand was used to immunoprecipitate. Bands corresponding to light-chain IgG were of equal intensity in all lanes, suggesting equal antibody loading. IP, Immunoprecipitation antibody; IB, immunoblotting antibody.