Journal: bioRxiv
Article Title: ATXN2 polyglutamine expansion impairs QKI-dependent alternative splicing and oligodendrocyte maintenance
doi: 10.1101/2025.08.08.669189
Figure Lengend Snippet: a, Graph depicting the dysregulation rates of commonly identified proteins in cerebellum and spinal cord samples from WT and KIN animals at terminal stage of 14 months old (r = 0.355, Pearson correlation). Pie charts depict the total number of commonly identified proteins (2999), all dysregulated proteins with >0.5 log 2 (fold change) difference in all directions (196), among which there were 91 proteins commonly upregulated (red), 84 proteins commonly downregulated (green), 12 proteins down in cerebellum, up in spinal cord (blue), and 9 proteins up in cerebellum, down in spinal cord (purple). b, Protein-protein interaction network and pathway enrichment analysis of the commonly downregulated proteins (with <70% abundance in KIN) showed a cluster of multiple myelin (in shades of purple) and axonal/synaptic proteins (in shades of blue and orange), with the term “Myelin sheath” appearing as the strongest dysregulation after the term “Cytoplasm”. c, Protein and transcript levels of neuronal compartment factors in WT and KIN cerebellum (Cb) at pre-onset (3 mo) and terminal (14 mo) stages, showing late onset downregulations for NEFH and NPTN, with an earlier downregulation of NEFL protein. ACTB was used as loading control, and normalizations against CALB1 were performed in order to test the neuropil affection versus Purkinje cell soma. All transcripts showed a significant downregulation at pre-onset stage with progressive reduction during disease course in KIN tissue. Tbp was used as housekeeping gene in qRT-PCR experiments. Each data point represents a single animal. d, Protein and transcript levels of myelin compartment factors in WT and KIN cerebellum at pre-onset (3 mo) and terminal (14 mo) stages, showing prominent downregulations for CNP, MBP and PLP1, with a massive reduction of MOG and MAG proteins. Slight yet significant reductions were observed in MBP and PLP1 levels at the pre-onset stage, accompanying a stronger downregulation of MAG. ACTB was used as loading control, and normalizations against CALB1 were performed in order to test the myelin affection versus Purkinje cell soma. Mbp and Plp1 transcripts showed a progressive downregulation throughout the disease course in KIN tissue, however Cnp , Mog and Mag transcripts interestingly showed a significant upregulation at the pre-onset stage which disappeared at the terminal stage. Tbp was used as housekeeping gene in qRT-PCR experiments. Each data point represents a single animal. Immunohistochemical assessment and quantification of MAG (e) and CNP (e) proteins in WT and KIN cerebellum at pre-onset stage confirms the immunoblot findings with a prominent reduction of MAG abundance and no change in CNP levels in KIN tissue. At least two sections were quantified from two WT and KIN animals. DAPI (blue) marks the nuclei in granular layer.
Article Snippet: The TaqMan Assays utilized for this study were: Actb (Mm02619580_g1), Aspa (Mm00480867_m1), Atxn2 (Mm01199894_m1), Calb1 (Mm00486647_m1), Cnp (Mm01306641_m1), Hapln1 (Mm00618325_m1), Hapln2 (Mm00480745_m1), Hapln3 (Mm00724203_m1), Hapln4 (Mm00625974_m1), Ina (Mm00840982_m1), Mag (Mm00487538_m1), Mal (Mm01339780_m1), Mbp (Mm01266402_m1), Mobp (Mm02745649_m1), Mog (Mm00447824_m1), Nat8l (Mm01217217_m1), Nefh (Mm01191456_m1), Nefl (Mm01315666_m1), Nefm (Mm00456201_m1), Nptn (Mm00485990_m1), Plp1 (Mm01297210_m1), Rtn4 (Mm00445861_m1), Tbp (Mm00446973_m1), Tuba4a (Mm00849767_s1).
Techniques: Control, Quantitative RT-PCR, Immunohistochemical staining, Western Blot
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