Figure 4.

FASN inhibition decreases sensory neuron growth in vitro. (A) Representative images of TUJ1-labeled sensory neurons cultured from L4 dorsal root ganglion (DRG), that were treated with vehicle or 1 um of platensimycin 1 h after plating, and then allowed to grow for 24 h in culture. (B) Quantification of (A) indicating the average neurite length for each neuron for each condition. Automated neurite tracing and length quantifications were performed using Nikon NIS-Elements (Version 4.60). The specific analysis code for digital reconstruction of neurites is available upon request (requires NIS-Elements and General Analysis https://www.microscope.healthcare.nikon.com/products/software/nis-elements) (n = 4 mice/condition; Unpaired t-test). (C) Embryonic DRG spot cultures axotomized at DIV7 after a 24 h pre-treatment with DMSO as a control, platensimycin or platensimycin combined with rosiglitazone. Cultures were fixed after 24 h and stained with SCG10. Scale Bar: 50 µm. (D) Quantification of (C) indicating the distance of regenerating axons measured from the injury site. (E,F) Heatmap of genes for known axonally-trafficked protein genes 1 and 3 days after SNI and SCI in whole DRG or fluorescence-activated cell sorted (FACS) ascending DRG sensory neurons from Thy1YFP16 mice. (SNI (n = 3 mice) and SCI (n = 4 mice) for C; n = 3 mice per group, except n = 4 mice for naïve and SCI 1d for D). (G) Schematic indicating the experimental design for the in vivo conditioning experiment. (H) Representative images of horizontal sections of the dorsal column labeled with SCG10 and dextran-labeled conditioned axons that was injected into the ipsilateral sciatic nerve. (I) Quantification of (H) indicating the ratio of mean SCG10 staining intensity between conditioned and unconditioned axons of ascending DRG sensory neurons (n = 3 mice; One-sample t-test and Wilcoxon signed rank test).