Figure 6.

Mature BDNF is more effective than proBDNF in eliciting TrkB activation. Secreted proBDNF-His or mature BDNF-His was purified by Ni-chromatography. A, TrkB-expressing PC12 cells were cultured in serum-free media for 6 h, followed by treatment with 1 nm mature BDNF, 1 nm proBDNF, or diluent control (None) as indicated for 10 min. Cell lysates were prepared and immunoprecipitated (IP) with anti-Trk antisera, followed by SDS-PAGE, and Western blotted (WB) with the indicated antisera. B, TrkB-expressing PC12 cells were treated, or not, with proBDNF-His, mature BDNF-His, or commercial mature BDNF in media containing 1% serum. Forty-eight hours later, the percentages of neurite-bearing cells were scored by an observer blinded to the treatment conditions. Error bars indicate the values obtained from two independent experiments.