JCS039024 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Addition of EGF to serum-starved cells induces redistribution of flotillin proteins to late endosomes/lysosomes. Flotillin-2 and the late endosome/lysosome marker CD63 were detected by indirect immunofluorescence in HeLa cells.

• Supplemental Figure S2 -

  Fig. S2. Expression of active forms of Lyn and Src does not alter the distribution of flotillin-1 or caveolin-1. Flotillin-1 and caveolin-1 were detected by indirect immunofluorescence in HeLa cells, transfected as shown.

• Supplemental Figure S3 -

  Fig. S3. Expression of active Fyn has no effect on transferrin uptake, but active Src blocks this process. (A) Transfection with SrcY527F-mCh blocks transferrin uptake. HeLa cells were labelled with FITC-transferrin for 20 minutes. (B) Quantification of total transferrin after uptake for 20 minutes in control cells (transfected with mCh alone), cells transfected with FynY531F-mCh, and cells transfected with SrcY527F-mCh. Mean fluorescence intensity of the transfected cells was normalized to that of surrounding untransfected cells. n=12, bars represent s.e.m.

• Movie 1 -

  Movie 1. Time-lapse total internal reflection images of the cell shown in Fig. 4A, acquired at 2Hz and played back at 10× real time. Red is Fyn-mRFP, green flotillin-1-GFP.

• Movie 2 -

  Movie 2. Time-lapse total internal reflection images of the cell shown in Fig. 4A, image was zoomed in as in Fig. 4B, acquired at 2 Hz and played back at 10× real time. Red is Fyn-mRFP, green flotillin-1-GFP.