Actin ac 74

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.

The following antibodies were used: SPOP (ab137537; Abcam, Cambridge, UK), GREB1 (ab72999; Abcam), Cyclin D1 (ab134175; Abcam), ERα (8644; Cell Signaling, Beverly, MA, USA), ubiquitin (6652-1; Epitomics, Burlingame, CA, USA), Myc (9E10; Sigma Aldrich, St. Louis, MO, USA), FLAG (M2; Sigma), actin (AC-74; Sigma) and HA (MM5-101R; Millipore, Darmstadt, Germany).

Antibodies for immunoblotting were from Sigma (actin, AC-74 used at 1:5,000 dilution and caspase-11, 17D9 used at 1:1,000 dilution), Cell Signaling (caspase-7, 9492 used at 1:1,000 dilution), Adipogen (caspase-1 p20, AG-20B-0042 used at 1:1,000 dilution), DSHB (tubulin, E7 used at 1:5,000 dilution) and Santa Cruz Biotechnology (ASC sc-22514-R used at 1:1,000 dilution). Propidium iodide was from Life Technologies and Lipofectamine2000 from Invitrogen. siRNAs for caspase-7 and caspase-11 were purchased from Santa Cruz and used at 40 pmol. zVAD-FMK and YVAD-FMK were from R&D systems.