JCS019729 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. NCAM is localized in EEA-1-positive and caveolin-1-positive endosomes. (A) B35 cells were transiently transfected with NCAM140-eGFP or NCAM180-eGFP constructs. Endocytosis was induced for 1 hour by application of an NCAM-specific antibody. Cells were fixed and either EEA-1 or caveolin-1 was visualized with Cy3-conjugated secondary antibodies. (B) Quantification of internalized NCAM in EEA-1-positive or caveolin-1-positive endosomes. Data are the mean ± s.e.m. of three independent experiments with at least 15 cells analysed in each experiment.

• Supplemental Figure S2 -

  Fig. S2. Densitometric analysis of NCAM140-expressing or NCAM180-expressing B35 cells. (A,B) B35 cells expressing either NCAM140 (A) or NCAM180 (B) were mock treated, treated with chloroquine at different concentrations or with 10 μM lactacystin for 4 hours. During the last hour of incubation an NCAM-specific antibody was added to induce NCAM endocytosis. Cells were lysed, 15 μg of total protein of each sample were loaded onto an SDS gel and subjected to western blot analysis using an NCAM-specific antibody. The experiment was carried out twice and the mock treated controls were considered 100%. All other values were related to the corresponding control.