Skin-derived fibroblasts (Isabelle Meyts, Laboratory of Inborn Errors of Immunity, KU Leuven, Leuven, Belgium) were obtained from a healthy individual (CTR) and a patient with p.W681X/p.W681X nonsense mutation in EIF2AK3 (PERK) suffering from Wolcott-Rallison syndrome. Fibroblasts were isolated from a freshly obtained skin biopsy, treated overnight with collagenase Type III (Stemcell, 400 CDU/ml), plated in T25 culture flask in serum-free conditions. After the treatment, the cells were further maintained in Dulbecco’s modified Eagle’s medium/Nutrient Mixture F-12 containing 4.5 g/l glucose, 0.11 g/l sodium pyruvate and 2 mM glutamine and supplemented with 100 units/ml penicillin, 100 μg/ml streptomycin and 10% FBS.
All cells were maintained in 5% CO2 at 37°C. Cells were routinely checked for mycoplasma contaminations using the Plasmotest kit (Invivogen) according to the manufacturer’s instructions.
All cells were maintained in 5% CO2 at 37°C. Cells were routinely checked for mycoplasma contaminations using the Plasmotest kit (Invivogen) according to the manufacturer’s instructions.
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