Drebrin

Antibodies to APP (6E10, Covance, Madison, WI, USA), cofilin (D3F9, Cell Signaling, Danvers, MA, USA), phospho-cofilin (77G2, Cell Signaling), actin (AC-74, Sigma Aldrich, St. Louis, MO, USA), tubulin (TU-02, Santa Cruz Biotechnology, Dallas, TX, USA), Aβ (D54D2, Cell Signaling), GFAP (Invitrogen), synapsin I (Invitrogen), PSD95 (Abcam, Cambridge, MA, USA), Drebrin (Abcam), microtubule-associated protein 2 (Millipore, Billerica, MA, USA), HRP-linked secondary antibodies (Jackson Immunochemicals, West Grove, PA, USA), and fluorescently labeled secondary antibodies (Invitrogen) were obtained from the indicated sources. The mouse monoclonal anti-RanBP9 antibody was a generous gift from Professor Elizabetta Bianchi (Pasteur Institute, France). Synthetic Aβ1-42 peptide was purchased from American Peptide (Sunnyvale, CA, USA). Aβ1-42 oligomers were prepared as previously characterized.^{21 (link)} Briefly, Aβ1-42 powder was dissolved in hexafluoro-2-propanol (HFIP) at 1 mM for 30 min at room temperature, aliquoted to eppendorf tubes, allowed to evaporate overnight in fume hood, and subjected to speed vacuum for 1 h to remove traces of HFIP or moisture. To prepare Aβ oligomers, Aβ1-42 film was then dissolved in dimethyl sulfoxide (5 mM), and F-12 cell culture medium (without phenol) was added to a final concentration of 100 μM Aβ1-42 and incubated at 4 °C for 24 h.

Antibodies to total tau (Tau A10, 1:1000 for Western blotting, 1:200 for cell/tissue staining; Santa Cruz Biotechnology, Dallas, TX, USA), PHF1 (kind gift from Dr. Peter Davies, 1:20, Albert Einstein College of Medicine), Tau-pS199/pS202 (1:1000 for western blotting, 1:200 for tissue staining; Invitrogen, Carlsbad, CA, USA), HT7 (1:200 for cell/tissue staining; Invitrogen, Carlsbad, CA, USA), GFAP (1:1000; Invitrogen, Carlsbad, CA, USA), synaptophysin (1:200; Invitrogen, Carlsbad, CA, USA), drebrin (1:400; Abcam, Cambridge, MA, USA), MAP2 (EMD Millipore, Billerica, MA, USA), cofilin (1:1000; Cell Signaling, Denvers, MA, USA) tubulin (1:1000; Millipore, Billerica, MA), detyrosinated tubulin (1:200; Millipore, Billerica, MA), RFP (1:200; Abcam, Cambridge, MA, USA) were used. cofilin m-RFP adenoviral constructs and adenoviruses (WT, S3A, and S3E) were kind gifts from Dr. James Bamburg, Colorado State University (Fort Collins)^{25 (link)}.

Cdk5 (Abcam, Cambridge, UK; clone 2G2), cytokeratin 5 (Abcam, EP1601Y), cytokeratin 8 (BioLegend, London, UK; clone 1E8), drebrin (GeneTex, Isleworth, UK; GTX11068), drebrin (Abcam 176318 & 178408), drebrin (Progen, Heidelberg, Germany; clone Mx823), EB1 (BD Transduction Laboratories, Oxford, UK; clone 5), EB3 (Millipore, Consett, UK; AB6033), p35 (Abcam, ab66064), GAPDH (GeneTex, clone GT239), GFP (Abcam, 6556), laminin (Sigma-Aldrich, Poole, UK; clone LAM-89), p35/25 (Cell Signaling, Hitchin, UK; C64B10), p63 (Abcam, ab735 clone BC4A4), pERK1/2 (Thr202/Tyr204) antibody (Cell Signaling; #9101), pS142-drebrin (Millipore, clone 3C14), tyrosinated α-tubulin (SeroTec, Kidlington, UK; clone YL 1/2), vimentin (Dako, Glostrup, Denmark; clone V9). Horseradish peroxidase-conjugated secondary antibodies were from Dako. Alexa-conjugated secondary antibodies and phalloidin were from Life Technologies, Warrington, UK. BTP2 (YM-58483) was from Cambridge Bioscience (Cambridge, UK; CAY13246). Recombinant human CXCL12 was from PeproTech (London, UK; 300-28A). Matrigel was from Corning (New York, USA; 354234) and fibronectin from Millipore.