Fig 1: Dnm1+/R237W mice display no gross abnormalities but altered protein expression.a Brains from 2 month-old Dnm1+/+ and Dnm1+/R237W mice were perfusion-fixed and 50 µm brain sections were stained with Nissl and a NeuN antibody to label gross neuronal architecture. Cortex (CTX), thalamus (THA), hypothalamus (HYP), amygdala (AMG) and hippocampus (HPC) are labelled, scale bar 1 mm. Lower panels represent zoom images of the hippocampus, scale bar 200 µm. b, c Lysates from primary cultures of hippocampal neurons prepared from either Dnm1+/+ and Dnm1+/R237W embryos were prepared and blotted for common presynaptic proteins and dynamin-1 interaction partners. b Representative blots display levels of Synaptotagmin-1 (Syt1), Amphiphysin-1 (Amph1), C-src, Endophilin, Syndapin and Eps15. c Quantification of protein levels normalised to Dnm1+/+ ± SEM (n = 3 independent cultures for all, all ns, two-sided Mann-Whitney test). d, e Workflow of quantitative proteomic analysis. Total protein content was cleaned onto SDS-PAGE gel before tryptic digestion. Proteins were analysed by high-resolution tandem MS, with significant differences revealed using a two-sided unpaired t test corrected for multiple comparisons. e Volcano plot displays 4237 quantified proteins, 39 which were depleted in Dnm1+/R237W (blue), while 151 were enriched (red). Dnm1 level is displayed in green. f STRING network analysis of up and down regulated proteins in Dnm1+/R237W synaptosomes. The resulting sub-complexes were subjected to MCL clustering at granulosity 4, which results in 18 clusters including 2 clusters with an average superior to 4 for the up-regulated proteins (proteasome core complex, ErbB signalling pathways) and six clusters including two clusters with an average superior to 4 for the down-regulated proteins (GPCR signalling pathway, dynactin complex). Source data are provided as a Source Data file.
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