Journal: Brain Pathology
Article Title: Serpin neuropathology in the P497S UBQLN2 mouse model of ALS/FTD
doi: 10.1111/bpa.12948
Figure Lengend Snippet: UBQLN2 and serpin proteins form aggregates in P497S animals in an age‐dependent manner. (A) Representative images of filter trap assays of cortical tissue from 8‐ to 32‐week‐old P497S, WT356, and Non‐Tg animals indicates a significant increase in aggregation of UBQLN2, Serpin A1, and Serpin I1 in 32‐week‐old, but not 8‐week‐old, animals. (B) Quantification of filter trap assays: * p < 0.05, ** p < 0.01, **** p < 0.0001. n = 3‐6 animals/age and genotype. (C and D) Immunoblots of equal portion of fractions following sequential extraction of cortical tissues from 8‐ (C) to 32‐week‐old (D) Non‐Tg, WT356, and P497S mice with increasingly harsh solubilizing buffers (low salt buffer, Triton X‐100 (TX‐100), sarkosyl (SARK), and urea). The GAPDH signal was virtually undetectable in the urea samples, and therefore, is not shown. (E) Quantification of the UBQLN2 and Serpin A1 signals in the urea fractions for 8‐ and 32‐week‐old animals. * p < 0.05, ** p < 0.01, **** p < 0.0001
Article Snippet: Rat anti‐Serpin A1a (#MAB7690, IB, IF), rat anti‐Serpin C1 (#MAB1287, IB, IF, both from R&D Systems, Minneapolis, MN), mouse anti‐UBQLN2 #NBP2‐25164, IB, IF, Novus Biologicals, Littleton, CO), mouse anti‐neuroserpin (#66997‐1, IB, IF, Proteintech Group, Inc, Rosemont, IL), mouse anti‐ubiquitin (sc‐8017, Santa Cruz Biotechnology, IB, Santa Cruz, CA), Rabbit anti‐GFAP (#PA1‐10019 IF, Thermo Fisher Scientific), rabbit anti‐IBA1 (#019‐19741, IF, FUJIFILM Wako Pure Chemicals, Corp, Richmond, VA), rabbit anti‐Serpin B1 (#PA576875, IB, IF, Thermo Fisher Scientific), rabbit anti‐Serpin C1/antithrombin III (#NBP2‐76966, IB, Novus Biologicals), rabbit anti‐LC3A/B (#4108, IF, Cell Signaling Technology, Danvers, MA, USA), rabbit anti‐Myc (UMY81, IB, home‐made against a peptide with an amino acid sequence of MEQKLISEEDLN), rabbit anti‐Neuroserpin (#ab33077, IB, IF, abcam, Eugene, OR), rabbit anti‐p97/VCP (UMY475, IB, home‐made), rabbit anti‐UBQLN2 (#23449‐1‐AP, IF Proteintech Group, Inc.), goat anti‐ChAT (#AB144P, IF, Millipore‐Sigma, Burlington, MA), and goat anti‐LAMP1 (sc‐8098, IF, Santa Cruz Biotechnology (Dallas, TX, USA).
Techniques: Western Blot