Figure 1.

Specificity of polyclonal anti-strumpellin antibodies and expression pattern of endogenous and expressed strumpellin. Panel A shows western blot of rat brain homogenate using polyclonal anti-strumpellin antibodies (lane 1) with several identified bands, including the expected size of 137 kD, corresponding to a full-length strumpellin protein. The specificity of this band was confirmed by RNAi experiments targeting this transcript. The 137- and 70-kD bands disappeared (lane 3), while control experiments after RNAi using nonsensical shRNA (lane 2) did not differ from the untreated brain homogenates (lane 1). Panel B shows immunocytochemistry experiments using the same RNAi construct, which almost completely eliminated strumpellin staining from the neuronal processes (arrows), but we observed a nonspecific background in neuronal bodies. Control experiments with nonsensical shRNA transfection did not change strumpellin staining (panel C). Scale bar= 10 μm.