Anti hsc70

DOPC, DOPS, and PEG-DSPE, were purchased from NOF Corp. Ltd. (Tokyo, Japan). DOTAP (chloride salt), NBD-DMPE, and rho-DMPE were purchased from Avanti Polar Lipids (Alabaster, AL, USA). Anti-Hsc70 (Anti-Hsc70 rat monoclonal antibody [1B5]), anti-HER2 (anti-ErB2), anti-CD9, anti-phosphorylated HER2 (p-Neu[Tyr1248]), phycoerythrin (PE)-conjugated mouse anti-human CD63, horseradish peroxidase (HRP)-conjugated goat-anti rabbit-IgG, and PE-conjugated mouse IgG1 were purchased from Enzo Life Sciences (Farmingdale, NY, USA), abcam (Cambridge, UK), Santa Cruz Biotechnology (Santa Cruz, CA, USA), Biolegend (San Diego, CA, USA), and BD Bioscience (San Jose, CA, USA), respectively. ECL Western Blotting Detection Reagent and PD SpinTrap G-25 were purchased from GE Healthcare Japan (Tokyo, Japan). Aldehyde/sulfate latex beads (4% w/v, 4 μm) and stain buffer (FBS) were purchased from Life Technologies (Carlsbad, CA, USA) and BD Bioscience (CA, USA), respectively. CFSE was obtained from Dojindo Molecular Technologies, Inc. (Kumamoto, Japan).

Anti-HA antibodies were used for immunofluorescence (3F10 rat monoclonal, Roche Applied Science) or immunoblotting (rabbit sc-805, Santa Cruz Biotechnology). Other antibodies from Santa Cruz Biotechnology included emerin (sc-15378), lamin A (sc-6215), lamin B (sc-6216), PDI (sc-20132), RTN1/2 (sc-23881), RTN2 (sc-16682), RTN3 (sc-33599), and RTN4 (sc-25660). Anti-turboGFP was purchased from Evrogen, anti-BiP and anti-calnexin from Abcam, anti-nuclear pore Mab(414) from Covance, and anti-Hsc70 from Enzo (SPA-757). The Sec61γ antibody was the gift of P. Lusk (Yale University). Fluorescently-labeled donkey anti-mouse, rabbit and goat immunoglobulin G (IgG; H+L) were purchased from Invitrogen (Carlsbad, CA) and used as secondary antibodies. Horseradish peroxidase-labeled donkey anti-mouse, rabbit or goat secondary antibodies were purchased from Jackson Immunoresearch and used in immunoblots. A B12-specific monoclonal antibody (clone 2D15) was developed in collaboration with Ab-Mart (Arlington, MA) against the epitope PTDTTHATHR in human DNAJB12. This antibody performed well for immunofluorescence, immunoblotting, and immunoprecipitation. Immunoblotting with this antibody showed that B12 shRNA caused loss of the endogenous B12 protein and that exogenous B12 was expressed at higher levels than the endogenous protein (Figure S1C).