Fig. 5. Analysis of the protein content in the slice incubation buffer after hyaluronidase treatment. Silver-stained polyacrylamide gels are shown after electrophoresis of supernatants from hippocampal slices (P6): C, control without addition of enzyme; Hy, after hyaluronidase treatment; ABC, after treatment with chondroitinase ABC; Hy/ABC, after treatment with both hyaluronidase and chondroitinase. (A) Protein bands in the range between 200 kDa and 400 kDa are shown. Note the presence of an additional band (*) of approximately 300 kDa after hyaluronidase (Hy) treatment of the slices. This band is not seen in control supernatants (C) or in supernatants from chondroitinase (ABC) treated slices, suggesting specific release of this protein by hyaluronidase treatment. Absence of this band in supernatant that was treated with both hyaluronidase and chondroitinase (Hy/ABC) suggests that the band represents a protein that is modified by chondroitinsulfate. (B) Protein bands below 140 kDa are shown. Note the presence of an additional band (*) of approx. 130 kDa after hyaluronidase (Hy) treatment of the slices. This band is not seen in control supernatants (C) or in supernatants from chondroitinase (ABC) treated slices, suggesting release of this protein only by hyaluronidase treatment of the slices. However, this band is unaffected by treatment of slices with both hyaluronidase and chondroitinase, suggesting it is not modified by chondroitinsulfate.