Western blotting
Cells were lysed in modified RIPA buffer (Tris HCl (50 mM), EDTA (1 mM),
Halt Protease Inhibitor cocktail (Thermofisher), pH7.4. Western Blot
Analysis Protein content was measured using the Novex® protein
separation kit (Thermofisher). Equal amounts of protein lysates were
separated by SDS-PAGE before blotting onto nitrocellulose membrane.
Membranes were probed with primary antibodies (see Table S2), detected
with horseradish peroxidase conjugated secondary antibodies and
visualised by enhanced chemiluminescence (GE Healthcare). Quantitative
signals were derived by densiometric analysis using ImageQuant™ TL on an
ImageQuant™ LAS 4000 Luminescent Image Analyzer (Fujifilm).