Western blotting has been described previously [70 (link)]. Antibodies used included: rabbit anti-TRKA (#06-574; 1:1000) and mouse anti-pY (#05-1050; 1:2000) from Millipore; rabbit anti-TRK (#4609; 1:500) and rabbit anti-phospho-TRKA (#9141; 1:1000) from Cell Signaling; mouse anti-V5 (#R960-25; 1:5000) from Invitrogen; mouse anti-GAPDH (#G8795; 1:5000) from Sigma-Aldrich.
N-linked glycosylation was inhibited from 6 h after transfection by adding tunicamycin (2 µg/ml; AppliChem) and cells were lysed 9 h later in RIPA buffer.
Immunoprecipitation, immunofluorescence and confocal microscopy were done as described previously [70 (link)].
N-linked glycosylation was inhibited from 6 h after transfection by adding tunicamycin (2 µg/ml; AppliChem) and cells were lysed 9 h later in RIPA buffer.
Immunoprecipitation, immunofluorescence and confocal microscopy were done as described previously [70 (link)].
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