Fig. 5: hnRNP R is associated with translation initiation complexes in motoneurons.
From: hnRNP R promotes O-GlcNAcylation of eIF4G and facilitates axonal protein synthesis
a Immunoblot analysis of hnRNP R co-immunoprecipitation by Y10b from motoneuron and NSC-34 cell lysate with or without RNase I treatment. Gapdh and eS5 were used as a negative and positive control, respectively. The immunoblot is representative of two biological replicates. b Schematic of a microfluidic chamber for compartmentalized motoneuron cultures allowing separation of the somatodendritic from axonal compartments. c Immunostaining of motoneurons cultured in a compartmentalized microfluidic chamber. Scale bars, 100 µm, and 100 µm (magnified areas). The images are representative of at least three biological replicates. d Absorbance profile of somatodendritic lysate subjected to sucrose density gradient ultracentrifugation and fractionation. Data are representative of two biological replicates. e Immunoblot analysis of hnRNP R and eIF2α in sucrose gradient fractions from somatodendritic and axonal lysates. For fractions 1 and 2 of the somatodendritic compartment, 10% were loaded for analysis. Data are representative of two biological replicates. f Quantification of 18S rRNA by qPCR analysis in sucrose gradient fractions from somatodendritic and axonal lysates. Data are representative of two biological replicates. g Immunoblot analysis of proteins co-immunoprecipitated by an anti-hnRNP R antibody from motoneuron lysate without or with Benzonase treatment. The immunoblot is representative of two biological replicates. h Representative images of hnRNP R-eIF2α PLA signal in cultured motoneurons at DIV 6 using anti-hnRNP R and anti-eIF2α antibodies. Scale bars, 10 µm, and 5 µm (magnified areas). The images are representative of two biological replicates. Source data are provided as a Source Data file.
