Neuronal Profilin Isoforms Are Addressed by Different Signalling Pathways
Figure 1
Specificity of the monoclonal antibodies raised against PFN1 (2C5) or PFN2a (4H5).
(A): Isoform-specific epitopes recognised by the monoclonal antibodies 2C5 and 4H5, as determined by pepscan analysis on overlapping 15mer amino acid sequences, as described [51]. Their respective location on the surface of PFN1 or PFN2a (blue) is indicated in the structural models designed by using Pymol software (DeLano Scientific LLC, Palo Alto, USA, version 0.98). (B): PFN isoform specific reactivity of 2C5 and 4H5. (B1): Immunoblot of 2C5 with purified recombinant mouse PFN1, mouse brain PFN2a and total extract of mouse spleen. (B2): Dot blot (left) and immunoblot (right) of 4H5 with recombinant mouse PFN1, mouse brain PFN2a and total brain extract. BSA: bovine serum albumin used as control. (C): Confocal images of immunofluorescence with 2C5 (upper panels) and 4H5 (lower panels) of C2C12 mouse myoblasts transfected with GFP-PFN2a. The antibody 2C5 reveals a typical fine diffuse cytoplasmic, but also a nuclear staining for PFN1, while the PFN2a specific antibody 4H5 labels only the GFP-PFN2a transfected cells (lower left) as part of a C2C12 cell population (DIC image; lower right). Filamentous actin was stained with FITC-phalloidin. Bars: 10 µm.