Hela cell line

The cerebellar granule (Cb) cell line derived from CD1 mice was a kind gift from Dr. Susan Cotman (MGH, Harvard Medical School, Boston, MA, USA) and maintained as described previously [23] (link). In short, the cells were grown in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with d-glucose, L-glutamine and pyruvate, with the addition of foetal calf serum, penicillin, streptomycin, Geneticin (Life Technologies, Darmstadt, Germany), and potassium chloride (Sigma Aldrich, Munich, Germany). The cells were maintained in a humidified incubator set to 33°C under 5% CO₂ atmosphere. The HeLa cell line (LGC Standards GmbH, Wesel, Germany) was grown in DMEM supplemented with glucose, foetal calf serum, penicillin and streptomycin in a humidified cell incubator set to 37°C under 5% CO₂.

The HeLa cell line was purchased from ATCC/LGC Promochem (Wesel, Germany) and cultured in RPMI (Sigma, cat. No. D8758, Deisenhofen, Germany); containing 10% Fetal Calf Serum (FCS, Biochrom GmbH, Cat. No. S0615, Berlin, Germany) and 1% penicillin/streptomycin (Sigma, cat. No. P433) and maintained in a humidified atmosphere of 5% CO₂ at 37°C. Cells were stably transfected with a pcDNA3.1 control plasmid (Invitrogen, Karlsruhe, Germany) or a plasmid allowing for the overexpression of wild-type (WT), a constitutively membrane-bound (Sdc-1-388), and a constitutively shed form (Sdc-1-392) of murine Sdc-1 in the vector pReceiver-M02 under control of the cytomegalovirus promoter (RZPD/ImaGenes, Berlin, Germany) as previously described (10 (link), 20 (link)). Stable clones were selected using 1 mg/ml G418. HeLa cells were cultured in RPMI-1640 medium containing 10% FCS, 1% penicillin/streptomycin and 600 mg/ml G418 in a humidified atmosphere of 5% CO₂ at 37°C. Successful transfections were confirmed by qPCR.