JCS02794 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Cells were transfected with YFP or dSH2-YFP. 24 hours post transfection cells were lysed and immunoprecipitation was performed using anti-GFP antibodies. IPs were run on a 4-12% gradient gel and the precipitated proteins were then transferred to nitrocellulose. Western blots were probed by antibodies (left). After stripping the blot they were re-probed with anti-GFP antibodies (right). Lane 1: whole cell lysate from cells expressing dSH2-YFP. Lane 2: IP GFP of cells expressing YFP. Lane 3: IP GFP of cells expressing dSH2-YFP. No vinculin was detected to co-precipitate in cells expressing dSH2-YFP (lane 3 left).

• Supplemental Figure 2 -

  Fig. S2. Presence of ‘hot spots’ when donor and acceptor fluorophores are switched between dSH2 and Paxillin.

• Supplemental Figure 3 -

  Fig. S3. Left two images: dSH2 fused to YFP and CFP in relative intensity color scale. Right image: FRET calculated without background subtraction or thresholding. Note the low FRET in the cytoplasm, suggesting that there is no self-FRET due to dimerization of the probe outside the FA area.