Titration of the lentivirus of sh-Nmnat2 in primary mouse DRG neurons. (A and B) Representative western blot image (A) and quantification (B) of NMNAT2 protein levels in primary mouse DRG neurons treated with the indicated amounts of sh-Nmnat2 lentivirus for 3 days. The NMNAT2 levels are normalized to Tuj1. (C and D) Representative phase contrast images (C) and quantification (D) of percentage of degenerated DRG axons 3 days after sh-Nmnat2 lentivirus infection. The concentration of the sh-Nmnat2 lentivirus is ∼2.38 × 10⁶ viral particles/μl. Infection of the DRG cultures with 40 μl of sh-Nmnat2 lentivirus for 3 days dramatically decreased NMNAT2 protein levels without inducing significant spontaneous axon degeneration. As such, this condition was used in the experiments in Fig. 2, D–F. Mean ± SEM. N = 4 in B and n = 7 DRG cultures per group from pooled results of three independent repeats in D. One-way ANOVA; ***P < 0.001; ns, not significant. Scale bar: 50 µm. Source data are available for this figure: SourceData FS3.