Oddyssey clx imaging system

GBM cells were lysed in an ice-cold buffer containing 0.2% DDM (N-Dodecyl-beta-Maltoside) and protease/phosphatase inhibitors (both from Sigma-Aldrich). The Pierce BCA Protein Assay kit (ThermoFisher Scientific) was used to determine the protein concentration in the DDM extracts. Cell debris was removed by centrifugation, and the lysates were incubated with an SDS-Loading buffer containing DTT (Sigma-Aldrich) at a final concentration of 16.6 mM. Samples were separated by SDS-PAGE, followed by transfer to nitrocellulose membranes (GE Healthcare Amersham, Chalfont Saint Giles, UK). The membranes were incubated with 8 µg/mL mouse anti-Tctex1 antibodies (Sigma-Aldrich) or 1 µg/mL Calnexin antibodies (Enzo Life Sciences, Lörrach, Germany) overnight at 4 °C, followed by IRDye^®-coupled secondary antibodies (LI-COR Biosciences, Bad Homburg, Germany) for 1h at room temperature. Signal detection was performed on a LI-COR Oddyssey^® CLx imaging system.