Protein disulfide isomerase pdi

Vero and QM5 cells were maintained at 37˚C and 5% CO₂ in medium 199 with Earle's salts supplemented with 5 or 10% fetal bovine serum (FBS), respectively_. The rabbit polyclonal anti-p14 and anti-14 ectodomain antisera were previously described (Corcoran and Duncan, 2004 (link); Top et al., 2005 (link)). Antibodies against actin (Sigma-Aldrich, St. Louis, MO), myc epitope tag (Sigma-Aldrich), PI4KIIIβ (BD Biosciences, Franklin Lakes, NJ), TGN46 (Serotec, Oxford, UK), protein disulfide isomerase (PDI; Enzo Life Sciences, Farmingdale, NY), horseradish peroxidase (HRP)–conjugated goat anti-rabbit (Jackson ImmunoResearch, West Grove, PA), Alexa Fluor 488–conjugated goat anti-mouse or donkey anti-sheep, Alexa Fluor 555–conjugated donkey anti-rabbit, and Alexa Fluor 647–conjugated goat anti-rabbit secondary antibodies (Life Technologies, Carlsbad, CA) were obtained from the indicated suppliers.

The chemicals used in this study were cycloheximide (CHX), MG132, 1,2-bis (o-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid (BAPTA) and 1,2-bis (o-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid acetoxymethyl ester (BAPTA-AM) (Sigma-Aldrich, St Louis, MO, USA); Nutlin-3 (TOCRIS, Avonmouth, Bristol, UK); Rhod-2-acetoxymethyl ester (Rhod-2-AM), Fluo-3-acetoxymethyl ester (Fluo-3-AM), MitoTracker-Red (MTR), MitoTracker-Green (MTG), calcein-acetoxymethyl ester (calcein-AM), ethidium homodimer-1 (EthD-1), and 4′,6-diamidino-2-phenylindole (DAPI) (Molecular Probe, Eugene, OR, USA); Ru360 (Calbiochem, Darmstadt, Germany); bortezomib, and carfilzomib (Selleckchem, Houston, TX, USA). The antibodies used in this study were anti-β-actin, ubiquitin, ATF4, mitochondrial Ca²⁺ uniporter (MCU), HDM2 and p53 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); CHOP/GADD153, GRP78, and mtHsp70 (Cell Signaling Technology, Danvers, MA, USA); cytochrome oxidase subunit II (COX II) (Invitrogen, Grand Island, NY, USA); protein disulfide isomerase (PDI) (Enzo Life Sciences, Farmingdale, NY, USA); rabbit IgG HRP, mouse IgG HRP and goat IgG HRP (Molecular Probe).