The Journal of Neuroscience, June 1, 2005, ():
ProBDNF Induces Neuronal Apoptosis via Activation of a Receptor Complex of p75NTR and Sortilin
J. Neurosci. Teng et al.
25: 5455
Supplemental Data
Files in this Data Supplement:
- supplemental material
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Supplementary Figure 1. p75NTR expressing smooth muscle cells were treated, or not (diluent), with purified proBDNF or mature BDNF at the indicated concentrations for 18 hrs. Cells were fixed and processed for TUNEL analysis and counterstained with DAPI. A minimum of 200 cells per condition (performed in triplicate) were evaluated by an observer blinded to condition. Vertical error bars indicate the averages from two independent experiments.
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Supplementary Figure 2. NGF partially rescues SCG neurons from proBDNF-induced death. Cultured rat SCG neurons, prepared as described (Fig. 7), were treated or not with 4 ng/ml proBDNF in the presence or absence of recombinant mature NGF (from 0.1 ng/ml to 10 ng/ml as indicated). Percentage ofSCG neurons death was determined under each treatment condition 48 hrs later. Vertical error bars indicate average values from duplicate cultures. Comparable results were obtained from two additional independent experiments.
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Supplementary Figure 3. Effects of proBDNF or proBDNF:sortilin complex on Schwann cells apoptosis. Rat Schwann cells were transfected with human sortilin using calcium phosphate, and treated with the indicated ligands (proBDNF and proBDNF:sortilin complexes were purified as noted in Fig. 8A). Sixteen hours later, cells were processed for double labeling by TUNEL and anti-sortilin immunofluorescence, followed by nuclear counterstaining with DAPI. TUNEL positivity in sortilin expressing cells was quantified by two observers blinded to condition. At least 1000 cells were scored per condition, and each condition was assessed in triplicate. Vertical error bars indicate the averages from two independent experiments.
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