Establishing NDP52 and p62 KO HeLa Cells

The NDP52 knock-out (KO) and p62 KO HeLa cells, as well as their corresponding control HeLa cells, which express NDP52 or p62, respectively, were kindly provided by Prof. Richard J. Youle (National Institutes of Health, Bethesda, Maryland, USA) (Lazarou et al., 2015 (link)). HeLa cells expressing the GFP-LC3 construct was kindly given by Drs. Aurore Rozières and Christophe Viret (CIRI, Centre International de Recherche en Infectiologie, Lyon, France). HeLa cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) with L-Glutamine (Gibco) supplemented with 10% fetal bovine serum (FBS) (Dutscher) and 1% antibiotic/antimycotic solution (Penicillin G/Streptomycin/Amphotericin B) (Cytiva). The intestinal epithelial cell line T84 (ATCC, CCL-248) was cultured in DMEM/F-12 Nutrient Mixture (Ham) (Gibco) supplemented with 10% FBS, 1% Glutamine (Gibco), 1% antibiotic/antimycotic solution (Penicillin G/Streptomycin/Amphotericin B) and 1% Hepes (Dutscher). All cells were maintained in an atmosphere with saturated humidity and containing 5% CO₂ at 37°C.