1. The use of fluorescence resonance energy transfer (FRET) to study the interaction between CD147 and MCT1 The principle of FRET MCT1 CD147 s s s s N- terminus C- Membrane CFP YFP CFP YFP OR N- terminus C- Energy transfer from excitation of the donor molecule (CFP) to the acceptor molecule (YFP) will occur only if the molecules are less than100Å apart. This causes quenching of donor fluorescence and excitation of the acceptor at its emission wavelength. Potential combinations of CFP- and YFP-tagged CD147 and MCT1

3. 0 2 4 6 8 10 12 460480500520540560580600 Emission wavelength nm intensity 0 2 4 6 8 10 12 14 460480500520540560580600 Emission wavelength nm intensity YFP laser excitation Co - transfection MCT1nCFP + CD147cYFP 0 5 10 15 20 25 30 460480500520540560580600 Emission wavelength nm intensity CFP laser excitation MCT1 CFP MCT1 s s s s CFP YFP CD147 CFP laser excitation Singletransfection CD147cYFP Singletransfection MCT1nCFP s s s s YFP CD147 No FRET FRET FRET occurs betweenMCT1nCFP andCD147cYFP but not between MCT1nCFP andCD147nYFP 0 5 10 15 20 25 460480500520540560580600 Emission wavelength nm intensity 0 10 20 30 460480500520540560580600 Emission wavelength nm intensity MCT1 CD147 s s s s CFP YFP Co-transfection with MCT1cCFP + CD147nYFP CFP laser excitation YFP laser excitation No FRET N

7. N N C N C out in out in C1 C2 Ca Cb N1 N2 Possible topology of MCT1/CD147 complex Monomer Dimer of 2 monomers