Fig. 2: Axonal protein synthesis is impaired in Hnrnpr−/− motoneurons.
From: hnRNP R promotes O-GlcNAcylation of eIF4G and facilitates axonal protein synthesis
a Immunostaining of DIV 6 puromycin-pulsed motoneurons cultured from Hnrnpr−/− and +/+ mice, with proximal and distal regions of the axon marked. Scale bars, 10 µm and 5 µm (magnified areas). b SuperPlots of puromycin immunosignals in the somata and proximal and distal axonal regions of Hnrnpr−/− and +/+ motoneurons. Statistical analysis was performed using an unpaired two-tailed Student’s t test. Data are mean ± s.d. of three biological replicates. Hnrnpr+/+, n = 69; Hnrnpr−/−, n = 60 motoneurons. c Schematic of a microfluidic chamber for compartmentalized motoneuron cultures allowing separation of the somatodendritic from axonal compartments. d Immunostaining of DIV 7 motoneurons cultured in a microfluidic chamber. Scale bar, 100 µm. The images are representative of at least three biological replicates. e Volcano plot of differential protein abundance in the axonal compared to the somatodendritic compartment of wildtype motoneurons cultured in microfluidic chambers from four biological replicates. Statistical analysis was performed using an unpaired two-sided Student’s t test. f Volcano plots of proteins differentially enriched in axonal and somatodendritic compartments of Hnrnpr−/− compared to +/+ motoneurons cultured in microfluidic chambers from four biological replicates. Statistical analysis was performed using an unpaired two-sided Student’s t test. g Overlap of proteins in (f) significantly (P < 0.05) deregulated in the somatodendritic and axonal compartments of hnRNP R knockout motoneurons. h Box plots highlighting differential enrichment of ribosomal proteins, translation factors, synaptic proteins and cytoskeletal components in axons of Hnrnpr−/− relative to +/+ motoneurons cultured in microfluidic chambers from four biological replicates. Statistical analysis was performed using Welch’s t test. i, Enrichment scores of protein subgroups in axons of Hnrnpr−/− relative to +/+ motoneurons. The size of the circles represents the number of proteins.
