Fig 6.

PGRN promotes survival of primary DRG neurons in the culture. (A) Immunofluorescence staining of primary DRG neurons transduced with indicated lentivirus particles and counterstained for neurofilament of 200 kD (red, NF200) and EGFP (green). (B) Representative life images of primary DRG cultures 48 hrs after transduction with indicated lentiviral particles. (C) Time courses of percentage survival of adult primary DRG neuron cultures (n = 4 cultures per treatment) after transduction with indicated viruses. Asterisks indicate significant differences between shGrn versus control virus (pLL-3.7), P < 0.05. (D) Numbers of surviving primary DRG neurons 96 hrs after NGF withdrawal from the culture medium, with or without recombinant human PGRN, as measured by counting the number of surviving neurons and expressed as percentage of baseline, taken immediately before transduction. At baseline images captured 298 ± 17 neurons per microscopic field. Data are means ± S.E.M. of triplicates. Asterisks indicate significant differences versus NGF-free/PGRN-free cultures, P < 0.05. ANOVA with subsequent Bonferroni t-tests.