Helam

HeLa, HeLaM, and HEK293T and COS7 cells were obtained from the American Type Culture Collection. Control. VAP-A/B double knock-out^{56 (link)} and parental HeLa cells were a gift from Prof Pietro De Camilli’s laboratory (Yale University School of Medicine, New Haven, CT, USA). Cells were cultured in DMEM with 4.5 g/l glucose (Lonza) containing 10% heat-inactivated FBS (Gibco) and 100 U/ml penicillin and 100 μg/ml streptomycin (Gibco) during experimental procedures. Human INPP5A 4 bp deletion KO cell line and a C631 human HAP1 parental control cell line was purchased from Horizon Discovery (HZGHC006619c008). Cells were routinely tested for Mycoplasma contamination and all tests were negative.

HeLaM, hTERT-RPE-1 and WT MEF cells were from American Type Culture Collection (ATCC, Manassas, VA, USA). Vimentin KO MEFs from a vimentin^-null mouse model were a kind gift from Dr Sara Koester (University of Gottingen) (Colucci-Guyon et al., 1994 (link)). Human skin fibroblasts derived from unaffected individuals and from individuals with GO were obtained from Dr May Tassabehji (University of Manchester, UK) and from the Cell Line and DNA Bank from Patients affected by Genetic Diseases (Genova, Italy). SW13 clone 1 (vim^+/+) and clone 2 (vim^−/−) were kind gifts from Victoria Allan (University of Manchester). All cells apart from hTERT-RPE-1 were cultured in GlutaMAX Dulbecco's modified Eagle medium (DMEM; Thermo Fisher Scientific, 31966021) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific, cat. no. A5256801), 50 U/ml penicillin and 50 µg/ml streptomycin. hTERT-RPE-1 cells were cultured in Ham's F12 and DMEM 1:1 (Thermo Fisher Scientific, 11320033) with the same supplements. Cells were cultured at 37°C in 5% CO₂. All cell lines were routinely tested for mycoplasma contamination.