Pen strep gln

Human lung adenocarcinoma A549 cells and Madin-Darby canine kidney (MDCK) cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Thermo Fisher Scientific, Ottawa, ON, Canada) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Grand Island, NY, USA) and 100 U/ml penicillin–100 μg/ml streptomycin–20 μg/ml glutamine (Pen/Strep/Gln; Wisent Bioproducts, St-Bruno, QC, Canada) at 37°C in a 5% CO₂ atmosphere. All cell lines were purchased from the American Type Culture Collection (ATCC). The generation of A549[EGFP-G3BP1] cells was described previously (22 (link)).

Unless specifically indicated, all chemicals were purchased from MilliporeSigma (Etobicoke, ON, Canada). PatA was a kind gift from Dr. Jerry Pelletier (McGill University, Montreal, QC, Canada), silvestrol was obtained from MedChem Express (Princeton, NJ, USA); 1 mM stocks of silvestrol and pateamine A were prepared in dimethyl sulfoxide (DMSO; MilliporeSigma, Etobicoke, ON, Canada) and stored at −80 °C. A549, Vero and Madin–Darby Canine Kidney (MDCK) cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Fisher Scientific, Ottawa, ON, Canada) supplemented with 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Grand Island, NY, USA) and 100 U/mL penicillin + 100 µg/mL streptomycin + 20 µg/mL-glutamine (Pen/Strep/Gln; Wisent Bioproducts, St-Bruno, QC, Canada) at 37 °C in 5% CO₂ atmosphere. All cell lines were purchased from the American Type Culture Collection (ATCC). PKR knock out A549 cells were generated by lentivirus-based CRISPR/Cas9 technique using guide RNA target sequence 5′-TAATACATACCGTCAGAAGC-3′ in lentiCRISPRv2 vector [21 (link)] and clonal selection.

Unless specifically indicated, all chemical reagents were purchased from Sigma. Pateamine A was a kind gift from Dr. Jerry Pelletier (McGill University, Montreal, QC, Canada). HeLa-TetOff cells were purchased from Clontech. A549, U2OS, Vero, and HeLa-TetOff cells were maintained in Dulbecco's modified Eagle's medium (DMEM; HiClone) supplemented with 10% fetal bovine serum (FBS, Life Technologies) and 100 U/ml penicillin+100 µg/ml streptomycin+20 µM L-glutamine (Pen/Strep/Gln; Wisent) at 37°C in 5% CO₂ atmosphere.