Rmnd5a nbp1 92337

Whole cell extracts were prepared as described [2 (link)] and resolved by SDS-PAGE (between 8% and 12%) and transferred to polyvinylidene difluoride (PVDF) membranes. Samples were analyzed with the following antibodies: HDAC6 (H-300, Santa Cruz, Santa Cruz, CA, USA, and PA5–11240, ThermoFischer Scientific), HA (HA-7, Sigma–Aldrich), Acetylated α-tubulin (6–11B-1, Santa Cruz, Santa Cruz, CA, USA), α-tubulin (T5168, Sigma–Aldrich), β-Actin (I-19, Santa Cruz, Santa Cruz, CA, USA), RanBPM (5 M, Bioacademia, Japan), Rmnd5A (NBP1–92337, Novus Biologicals) and muskelin (C-12, Santa Cruz, Santa Cruz, CA, USA). The blots were developed using Clarity ECL Western Blotting Substrate (BioRad, Hercules, CA). Quantifications were done using Image Lab (BioRad, Hercules, CA) and ImageJ software. Co-immunoprecipitation experiments were performed in 0.25% NP-40 and 100 mM KCl lysis buffer and were carried out overnight at 4 °C with antibodies to HA (HA-7, Sigma–Aldrich), OctA-Probe (D-8 Santa Cruz, Santa Cruz, CA, USA), HDAC6 (D-11 Santa Cruz, Santa Cruz, CA, USA) and RanBPM (F1 Santa Cruz, Santa Cruz, CA, USA). Immunoprecipitates were isolated with PureProteome Protein G Magnetic Beads (EMD Millipore, Billerica, Massachusetts) or Dynabeads Protein G (Invitrogen, Life Technologies, Burlington ON, Canada).

Whole cell extracts were prepared as described^{49 (link)} and resolved using 8% or 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Gels were transferred to polyvinylidene fluoride membranes, blocked in 5% milk and hybridized with the following antibodies: ARMC8 (E-1, sc-365307; Santa Cruz Biotechnology, Santa Cruz, CA, USA); FLAG (M2, F1804, Sigma-Aldrich, St. Louis, MO, USA); HA (HA-7, H3663 Sigma-Aldrich); MAEA (AF7288, R&D Systems, Minneapolis, MN, USA); Muskelin (C-12, sc-398956, Santa Cruz Biotechnology); RanBPM (5M, 71-001, Bioacademia, Japan); RMND5A (NBP1-92337, Novus Biologicals, Littleton, CO, USA); TWA1 (NBP1-32596, Novus Biologicals); Vinculin (E1E9V, Cell Signaling Technology, Danvers, MA, USA); WDR26 (ab85962, Abcam, Cambridge, UK); β-actin (A5441, Sigma-Aldrich) and His (MAB050, R&D Systems). Blots were developed using Clarity Western ECL Substrate (BioRad, Hercules, CA, USA) and imaged using a ChemiDoc MP (BioRad). Analysis and quantifications were done using Image Lab (BioRad).