Fig. 3

PGRN processing by cathepsins. a Cathepsin- and PGRN-deficient MEF cells were labeled with [³⁵S]-methionine and [³⁵S]-cysteine for 24 h and the cell lysates were then immunoprecipitated with rabbit anti-PGRN antibodies and separated by 16% Tricine-SDS PAGE. PGRN and PGRN-derived peptides were visualized by autography. * indicates non-specific bands. b Quantification of PGRN and PGRN-derived peptides in (a). 10 kDa PGRN-derived peptides were normalized with full-length PGRN signals in each group. Data is presented as means ± s.e.m. n = 3, * P,0.05; ***P < 0.001, ns, not significant, one-way ANOVA, Tukey’s Multiple Comparison Test. c Recombinant cathepsin D was incubated with recombinant human PGRN in acidic buffer for 16 h. Proteins were separated on a Bis-Tris gel and blotted with goat anti-human PGRN antibodies. d Recombinant cathepsin B and L were incubated with recombinant human PGRN, as indicated, in acidic buffer for 4 h. Proteins were separated on a Bis-Tris gel and blotted with goat anti-human PGRN antibodies